Excitatory glycine receptors and methods

ABSTRACT

The invention provides isolated NR3B polypeptides, functional fragments and peptides, encoding nucleic acid molecules and polynucleotides, and specific antibodies. Also provided are excitatory glycine receptors, containing either NR3B or NR3A polypeptides. Further provided are methods for detecting excitatory glycine receptor ligands, agonists and antagonists. The invention also provides related diagnostic and therapeutic methods.

[0001] This invention was made with United States Government support under grant numbers PO1 HD29587 and RO1 EY05477 awarded by the National Institutes of Health. The U.S. Government has certain rights in this invention.

BACKGROUND OF THE INVENTION

[0002] 1. Field of the Invention

[0003] This invention relates generally to the fields of neurobiology and medicine and, more specifically, to the field of ionotropic receptors. 2. Background Information

[0004] Ionotropic glutamate receptors activate ligand-gated cation channels that mediate the predominant component of excitatory neurotransmission in the central nervous system (CNS). These receptors have been classified based on their preference for the glutamate-like agonists (RS)-2-amino-3-(3-hydroxy-5-methyl-4-isoxazolyl)propionic acid (AMPA), kainate (KA), and N-methyl-D-aspartate (NMDA). All three glutamate receptor subtypes are heteromultimeric complexes, and many of the subunits that comprise them have been identified and characterized. To date, four AMPA receptor subunits (GluR1-4), five KA receptor subunits (GluR5-7, KA1, and KA2), and six NMDA receptor subunits (NR1, NR2A-2D and NR3A) have been reported.

[0005] The NMDA receptor (NMDAR) has unique properties that distinguish it from the other glutamate receptor subtypes. First, the activation of NMDAR requires the presence of dual agonists, glutamate (or NMDA) and glycine. In addition, activation of these receptors is regulated by Mg²⁺ in a voltage-dependent manner (i.e., the NMDAR is blocked at resting membrane potential and activated when depolarized). Most importantly, the NMDAR is extremely permeable to Ca²⁺, a key regulator of cell function. These unique properties allow NMDARs to play a critical role in development of the nervous system, synaptic plasticity, memory, and other physiological processes in the CNS. However, excessive stimulation of NMDARs has also been implicated in many pathological conditions including stroke, ischemia, head and spinal trauma, headache, epilepsy, neuropathic pain syndromes including diabetic neuropathy, glaucoma, depression and anxiety, drug addiction/withdrawal/tolerance, and in chronic neurodegenerative states, such as Alzheimer's disease, Huntington's disease, HIV-associated dementia, Parkinson's disease, multiple sclerosis, and amyotrophic lateral sclerosis (ALS).

[0006] The molecular cloning and functional analysis of expressed NR1, NR2A-D, and NR3A subunits, coupled with the examination of their temporal and spatial expression patterns in vivo, has led to significant advances in our understanding of NMDAR function at the molecular level. However, the identification of these six subunits alone has failed to explain the observed diversity in NMDAR function, particularly in motor neurons.

[0007] Thus, there exists a need to identify and characterize additional NMDAR subunits, and to characterize the function of additional NMDA receptors. There also exists a need to provide screening assays that identify compounds that modulate the function of additional NMDA receptors. Such compounds can be used to treat pathological conditions in which inappropriate NMDA receptor activation, or inappropriate responses to glycine or glutamate, are involved. The present invention satisfies these needs and provides related advantages as well.

SUMMARY OF THE INVENTION

[0008] The invention provides isolated nucleic acid molecules encoding N-methyl-D-aspartate (NMDA) receptor type 3B (NR3B) polypeptides, including human, rat and mouse NR3B polypeptides.

[0009] Also provided are vectors and cells containing isolated nucleic acid molecules encoding NR3B polypeptides.

[0010] The invention also provides a method of producing an NR3B polypeptide by expressing a nucleic acid molecule encoding an NR3B polypeptide in vitro or in a cell under conditions suitable for expression of the polypeptide.

[0011] Further provided are isolated NR3B nucleic acid molecules encoding functional fragments of an NR3B polypeptide, including functional fragments that bind glycine.

[0012] The invention also provides an isolated NR3B polynucleotide containing at least 17 contiguous nucleotides from a human, rat or mouse NR3B nucleic acid molecule.

[0013] Also provided is a method for detecting a nucleic acid molecule encoding a NR3B polypeptide in a sample, by contacting the sample with one or more NR3B polynucleotides, and detecting specific hybridization to the polynucleotide, thereby detecting a nucleic acid molecule encoding an NR3B polypeptide in said sample.

[0014] The invention further provides isolated NR3B polypeptides, including human, rat and mouse NR3B polypeptides.

[0015] Also provided are functional fragments of NR3B polypeptides, including functional fragments that bind glycine.

[0016] The invention also provides isolated NR3B peptides, containing at least 8 contiguous residues of an NR3B polypeptide.

[0017] Further provided is an isolated antibody or antigen binding fragment thereof, which specifically binds an isolated NR3B polypeptide.

[0018] Also provided is a method of detecting an NR3B polypeptide in a sample, by contacting the sample with an antibody which specifically binds an NR3B polypeptide, and detecting the presence of specific binding of the antibody to the sample, thereby detecting an NR3B polypeptide in the sample.

[0019] The invention also provides methods of detecting an NR3B ligand, by contacting an NR3B polypeptide or functional fragment with one or more candidate compounds under conditions suitable for detecting binding to the polypeptide, and detecting a candidate compound that binds the polypeptide, wherein such a compound is characterized as an NR3B ligand.

[0020] Further provided is a composition containing an isolated excitatory glycine receptor. In one embodiment, the excitatory glycine receptor contains and NR3B polypeptide and an NR1 polypeptide. In another embodiment, the excitatory glycine receptor contains and NR3A polypeptide and an NR1 polypeptide. Optionally, the receptor further contains an NR2A, NR2B, NR2C or NR2D polypeptide.

[0021] The invention also provides a method of detecting an excitatory glycine receptor ligand, by contacting an excitatory glycine receptor with one or more candidate compounds under conditions suitable for detecting binding to said receptor, and detecting a candidate compound that binds said receptor, wherein such a compound is characterized as an excitatory glycine receptor ligand.

[0022] Also provided is a method of detecting an excitatory glycine receptor agonist or antagonist, by contacting an excitatory glycine receptor with one or more candidate compounds under conditions suitable for detecting receptor activation, and detecting a candidate compound that alters receptor activation, wherein such a compound is characterized as an excitatory glycine receptor agonist or antagonist.

[0023] Further provided is a method of modulating a cellular response to glycine or glutamate, by introducing a nucleic acid molecule encoding an NR3B polypeptide or functional fragment into a cell, and expressing the NR3B polypeptide or functional fragment encoded by said nucleic acid molecule in said cell, whereby expression of the polypeptide or functional fragment modulates a cellular response to glycine or glutamate.

[0024] The invention further provides a method of modulating a cellular response to glycine or glutamate, by introducing an antisense nucleic acid molecule, a ribozyme molecule or a small interfering RNA (siRNA) molecule into the cell, wherein the molecule hybridizes to an NR3B nucleic acid molecule and prevents translation of the encoded NR3B polypeptide.

BRIEF DESCRIPTION OF THE DRAWINGS

[0025]FIG. 1 shows an alignment of the deduced amino acid sequence of the ionotropic glutamate receptor subunits from rat, designated NR1 (SEQ ID NO:14), NR2A (SEQ ID NO:15), NR3B B4 (SEQ ID NO:4) and NR3A (SEQ ID NO:16). Sequences were aligned using ClustalW and the BLOSUM series protein scoring matrix. Exact matches are boxed and shaded; conservative substitutions are boxed (no shading). Predicted signal peptide cleavage sites are indicated by vertical lines. Membrane regions (M1-M4) are indicated by horizontal lines. Asterisks indicate the positions of amino acid residues in NR1 and NR2A which have been shown to be required for glycine and glutamate binding, respectively. An arrow marks the positon of the conserved asparagine residue in NR1 and NR2A-D.

[0026]FIG. 2 shows the distribution of the NR3B subunit in the CNS by in situ hybridization. NR3B probes were labeled using isotopic (³³P)- (a, b) and non-isotopic (digoxigenin) (c, d) methods. Bar represents 6 mm for panel a, 3 mm for b, 50 mm for c, and 1 mm for d.

[0027]FIGS. 2a and 2 b show that positive signals (arrows) were detected only by probes derived from antisense (AS) sequences, but not with sense (S) probes in adult rat tissue. Strong NR3B signals were observed in facial and trigeminal nuclei of the brainstem and in the ventral horn of the spinal cord.

[0028]FIG. 2c shows NR3B-positive cells viewed under high magnification (400×, top panel). These cells resemble motor neurons retrogradely labeled by injection of a fluorescent dye (granular blue) into leg muscles (bottom panel).

[0029]FIG. 2d shows the distribution of the NR3B subunit in the lumbar spinal cord of rats at different ages. NR3B signals developed postnatally, appearing as early as P2, reached a peak around P14, and remained elevated in the adult. The positive cells are large and are located in layer VIII and IX, suggesting that they are motor neurons. Arrows pointing to the labeled motor neurons are placed only on the right side of the spinal cord.

[0030]FIG. 3 shows pharmacological characterization of NR1/NR3B receptors in Xenopus oocytes. Data are representative of recordings from 1-9 oocytes in each case. Currents were recorded in 1.5 mM Ba²⁺ Ringers' solution from oocytes injected with 2 ng of NR1 cRNA and 12 ng of NR3B cRNA.

[0031]FIG. 3a shows a dose-response of glycine-evoked currents from NR1/NR3B receptors. Apparent inhibition at glycine concentrations greater than 10 μM may result from fast desensitization kinetics not readily resolved by the two-voltage electrode recording system. Inset: Significantly slower time course of NMDA-induced desensitization of NR1/NR2A currents (100 μM NMDA plus 10 μM glycine). Rebound or resensitization of NR1/NR3B receptors at high glycine concentrations upon agonist washout may result from slower clearance of glycine from the oocyte-vitelline membrane space.

[0032]FIG. 3b _(1,3) show that D-serine, an NR1/NR2 receptor co-agonist, evokes small NR1/NR3B currents alone but inhibits glycine-evoked responses in a dose-dependent fashion. FIG. 3b ₂ shows that application of other NR1 glycine site agonists alone, including D-alanine, the cyclopropyl analogue ACPC or D-cycloserine (at concentrations sufficient to produce maximal or near maximal co-agonist activity at classical NMDARs) does not evoke detectable NR1/NR3B currents, but evokes NR1/NR2A currents when co-applied with 100 μM NMDA (inset). FIG. 3b ₃ shows that co-application of 10 μM glycine with maximally effective doses of D-serine, D-alanine, ACPC or D- cycloserine at classical NMDARs produces relative inhibition of glycine-evoked NR/NR3B currents.

[0033]FIG. 3c shows that NMDA (100 μM) does not potentiate 10 μM glycine-evoked currents.

[0034]FIG. 3d ₁₋₂ shows that kainate (100 μM) potentiates NR1/NR3B currents evoked by 5 μM glycine, while L-cysteine (50 μm), L -aspartate (100 μM) or glutamate (100 μM, not shown) have little effect or only slightly potentiate the current.

[0035]FIG. 3e ₁ shows that Mg²⁺ weakly inhibits glycine-evoked NR1/NR3B currents in a dose-dependent fashion, but much less potently than receptors containing NR2 subunits. Marked rebound or resensitization occurs upon Mg²⁺ washout. Inset: Mg²⁺ inhibition of NR1/NR2A receptors (bottom bar: 100 μM NMDA plus 10 μM glycine; top bar: 100 μM NMDA, 10 μM glycine, 0.1 mM Mg²⁺).

[0036]FIG. 3e ₂ shows that MK801 does not block and in fact appears to slightly potentiate glycine-evoked NR1/NR3B currents. Inset: 10 μM MK801 produces complete blockade of NR1/NR2A currents evoked by 100 μM NMDA plus 10 μM glycine.

[0037]FIG. 3e ₃ shows that Memantine (12 μM) only partially blocks glycine-evoked NR1/NR3B currents but completely blocks NR1/NR2A currents evoked by 100 μM NMDA plus 10 μM glycine (inset).

[0038]FIG. 3f ₁ shows that 10 μM D-aminophosphonovaleric acid (APV), a classical competitive NMDAR antagonist, has little effect on NR1/NR3B currents but blocks NR1/NR2A currents evoked by 100 μM NMDA plus 10 μM glycine (inset).

[0039]FIG. 3f ₂ shows that 5,7-dichlorokynurenic acid, an NR1 glycine site antagonist, completely inhibits NR1/NR3B currents evoked by glycine, while L-strychnine, an inhibitory glycine receptor antagonist, has no effect.

[0040]FIG. 4 shows whole-cell currents recorded from oocytes injected with NR1/NR3B (1:12) cRNA.

[0041]FIG. 4a shows currents recorded at a holding potential of −60 mV in response to application of 10 μM glycine. Application of 0.5 mM Mg²⁺ had no effect, but 100 mM N-methyl-D-gluconate (NMDG) completely blocked the current. The NMDA antagonist APV (40 μM) did not block the glycine-induced current, and actually potentiated the current.

[0042]FIG. 4b shows glycine-induced currents recorded at different membrane potentials.

[0043]FIG. 4c shows currents recorded in response to a voltage ramp from −60 to +80 mV (140 mV/sec). The currents reversed around −10 mV. In the presence of NMDG the current did not reverse. No current was carried by 100 mM Ca²⁺, suggesting that channels composed of NR3B subunits are not permeable to Ca²⁺ (not shown). These findings suggest that NR1/NR3B receptor-operated channels are permeable to monovalent cations, but not appreciably to Ca²⁺.

[0044]FIG. 5 shows single-channel recordings from outside-out patches obtained from oocytes injected with NR1/NR3B (1:12) cRNA.

[0045]FIG. 5a shows current recordings at a holding potential of −60 mV. Application of glycine (10 μM) activated single-channel currents, and Mg²⁺ (0.5 mM) had no significant effect on these currents. Single-channel currents are shown at higher time resolution below. The single-channel currents display a main conductance state (2.3 pA) and a sub conductance state (0.7 pA) in the presence of 2 mM Ba ²⁺ (the zero current level is shown as a dotted line).

[0046]FIG. 5b shows all point amplitude histograms of single channel currents of NR1/NR3B receptors activated by glycine.

[0047]FIG. 5c shows single-channel currents recorded at different membrane potentials.

[0048]FIG. 5d shows single channel current-voltage relationship of the main conductance (squares) and sub conductance (circle) states.

[0049]FIG. 6 shows the effect of NMDA receptor agonists and competitive antagonists on glycine-evoked NR1/NR3A currents

[0050]FIG. 6a shows that various concentration of glycine evoked desensitizing inward currents in oocytes injected with NR1 and NR3A subunits and held at −80 mV.

[0051]FIG. 6b shows that glycine-evoked currents were effectively blocked by D-serine, which in previously described NMDA receptors is normally a co-agonist binding to the NR1 subunit.

[0052]FIG. 6c shows that glycine-evoked currents under these conditions were only slightly blocked by APV, a competitive antagonist for previously described NMDA receptors.

[0053]FIG. 6d shows that 5, 7-di-Cl-Kynurenate, an antagonist that binds to the NR1 subunit, effectively blocked the glycine-evoked current of NR1/NR3A receptors.

[0054]FIG. 6e shows that the inhibitory glycine receptor blocker, strychnine, had no effect on glycine-evoked currents of NR1/NR3A receptors.

[0055]FIG. 7 shows the effect of open-channel blockers on glycine-evoked currents of NR1/NR3A receptors.

[0056]FIG. 7a shows that Mg²⁺ displays no inhibitory effect on NR1/NR3A receptors unlike its inhibitory effect on NMDA receptors containing NR2(A-D) subunits. At low glycine levels, Mg²⁺ actually potentiates glycine-evoked currents of NR1/NR3A receptors. At higher glycine levels, Mg²⁺ potentiates peak currents without affecting the steady state current. Inset: Mg²⁺ inhibition of NR1/NR2A receptors (Top bar: 100 μM NMDA plus 10 μM glycine; bottom bar: 100 μM NMDA, 10 μM glycine, 0.1 mM Mg²⁺).

[0057]FIG. 7b shows that MK801 potentiates glycine-evoked currents at both low (2.5 μM) and high (10 μM) glycine. Currents were recorded in 1.5 mM Ba²⁺ Ringers' solution.

[0058]FIG. 8 shows the deduced amino acid sequences of rat NR3B B4 (SEQ ID NO:4) and rat NR3B A2 (SEQ ID NO:2) and their alignment with the predicted sequences of mouse NR3B (SEQ ID NO:8) and human NR3B (SEQ ID NO:6). Sequences were aligned using ClustalW and the BLOSUM series protein scoring matrix. Exact matches are boxed and shaded; conservative substitutions are boxed (no shading). Gaps (−) were inserted to maximize homology. Thick horizontal lines indicate the positions of the predicted signal peptide and membrane regions (M1-M4). Dotted horizontal lines indicate the positions of the S1 and S2 ligand binding domains.

[0059]FIG. 9 shows an alignment of regions of ionotropic glutamate receptor subunits NR1, NR2A, NR2B, NR2C, NR2D, NR3A, NR3B and GluR2. Top: Residues of the S1 and S2 regions considered to be important for glutamate binding are indicated by *. Residues of the S1 and S2 regions considered to be important for glycine binding are indicated by ^ . Bottom: Residues of the channel lumen that are accessible from either one or both sides of the channel are boxed.

[0060]FIG. 10 shows the design of an NR3B targeting vector. The DNA fragment containing mouse NR3B exons 2-10 (˜7.6 kb) was replaced by a fragment containing the neomycin resistant (Neo^(r)) gene- (˜2 kb). The 5′- (˜3.6 kb) and 3′-(˜3.2 kb) arms used for homologous recombination are indicated by the thicker lines. The targeting DNA fragment was inserted into a pGTN29 vector.

[0061]FIG. 11 shows the predicted human NR3A cDNA sequence.

[0062]FIG. 12 shows the predicted human NR3A amino acid sequence.

DETAILED DESCRIPTION OF THE INVENTION

[0063] The present invention relates to the cloning and characterization of a seventh NMDAR subunit, designated herein NR3B. The present invention also relates to the determination that receptors containing NR3B, or receptors containing the previously identified NMDAR subunit NR3A, display strikingly distinctive properties from all previously characterized NMDARs. The invention provides molecules and methods that can be used to prevent or ameliorate conditions in which inappropriate NMDA receptor activation, or inappropriate responses to glycine or glutamate, are involved.

[0064] The invention provides an isolated nucleic acid molecule encoding a NR3B polypeptide. As used herein, the term “NR3B polypeptide” refers to a polypeptide that retains at least one biological activity characteristic of the naturally occurring mammalian NR3B polypeptides designated herein SEQ ID NOS:2, 4, 6 or 8. As disclosed herein, an exemplary biological activity characteristic of NR3B is the ability to form a subunit of an excitatory glycine receptor. An “excitatory glycine receptor” can be characterized as a receptor that responds to micromolar concentrations of glycine with a cation current. An excitatory glycine receptor can further be characterized by exhibiting any or all of the following properties: little or no response to NMDA or glutamate; little or no response to certain NR1 glycine site agonists, such as D-alanine, ACPC or D-cycloserine; inhibition of current in response to D-serine; inhibition of current in response to 5,7-dichlorokynuric acid; lack of inhibition of current in response to L-strychnine; lack of substantial inhibitory response to Mg²⁺, MK801 or memantine; enhancement of glycine-invoked current by ≧40 μM APV; relatively Ca²⁺-impermeable.

[0065] A further exemplary biological activity characteristic of NR3B is the ability to oligomerize with an NR1 subunit, and possibly further with both an NR1 and an NR2 subunit.

[0066] Yet another exemplary biological activity characteristic of NR3B is the ability to bind glycine with high affinity. The skilled person can determine other biological activities characteristic of an NR3B polypeptide designated herein SEQ ID NOS:2, 4, 6 or 8.

[0067] Isolated nucleic acid molecules encoding NR3B polypeptides can be used, for example, as templates for the recombinant expression of NR3B subunits (the uses of which are described in more detail below); as probes to detect NR3B-encoding nucleic acid molecules in samples; in in vivo and ex vivo gene therapy applications in which modulation of NR3B expression is desired; and in other therapeutic, diagnostic, screening and research applications known to those skilled in the art.

[0068] The term “isolated,” in reference to an invention nucleic acid molecule or polypeptide is intended to mean that the molecule is substantially removed or separated from components with which it is naturally associated, or is otherwise modified by the hand of man, thereby excluding nucleic acid and polypeptide molecules as they exist in nature. An isolated molecule can be in any form, such as in a buffered solution, a suspension, a lyophilized powder, attached to a solid support (e.g. as a component of an array, or on a filter or column), or in a cell or cell extract.

[0069] The term “nucleic acid molecule,” as used herein, refers to a polynucleotide of natural or synthetic origin. A nucleic acid molecule can be single- or double-stranded genomic DNA, cDNA or RNA, and can represent a sense strand, an antisense strand, or both. Accordingly, a designated sequence identifier, unless specified otherwise, is intended to refer to the single-stranded molecule having the recited sequence, the single-stranded complement of the recited sequence, or a double stranded (or partially double-stranded) molecule in which one strand has the recited sequence.

[0070] A nucleic acid molecule can optionally include one or more non-native nucleotides, having, for example, modifications to the base, the sugar, or the phosphate portion, or having a modified phosphodiester linkage. Such modifications can be advantageous in increasing the stability of the nucleic acid molecule. Furthermore, a nucleic acid molecule can include, for example, a detectable moiety, such as a radiolabel, a fluorochrome, a ferromagnetic substance, a luminescent tag or a detectable binding agent such as biotin. Such modifications can be advantageous in applications where detection of a hybridizing nucleic acid molecule is desired.

[0071] An isolated nucleic acid molecule encoding a NR3B polypeptide can encode SEQ ID NO:6, or encode a polypeptide having at least 60% identity to SEQ ID NO:6, such as at least 70%, 80%, 85%, 90%, 95%, 97%, 99% or greater identity to SEQ ID NO:6. Identity of any two nucleic acid or amino acid sequences can be determined by those skilled in the art based, for example, on known computer alignments such as BLAST 2.0, ClustalW and the like, which can be adjusted manually, if appropriate, to insert gaps to optimize the alignment according to standard practice in the art.

[0072] An isolated nucleic acid molecule encoding a NR3B polypeptide with at least 60% identity to SEQ ID NO:6 can encode a naturally occurring or a non-naturally occurring amino acid sequence. SEQ ID NO:6 represents the predicted amino acid sequence of a naturally occurring human NR3B polypeptide.

[0073] The skilled person will appreciate from the alignment shown in FIG. 8 that the C-terminus of SEQ ID NO:6 differs somewhat from the rat and mouse orthologs. Based on these observed differences, it is contemplated that a naturally occurring human NR3B polypeptide can contain additional sequence corresponding to one or more of the gaps where SEQ ID NO:6 does not apparently align with SEQ ID NOS: 2, 4 and 8, or no sequence in these positions.

[0074] For example, it is contemplated that a naturally occurring human NR3B polypeptide can contain several additional amino acids (e.g. 1-20 additional amino acids, such as 11 additional amino acids), or no amino acids, between the sequence “PPEGS” and “KEETA” of SEQ ID NO:6. Such additional sequence can be identical to, substantially similar to, or different from, the sequence QQERAEQERSGP (portion of SEQ ID NO:4) or the sequence QQERAEQECRGP (portion of SEQ ID NO:8). Likewise, it is contemplated that a naturally occurring human NR3B polypeptide can contain several additional amino acids (e.g. 1-20 additional amino acids, such as 8 additional amino acids), or no amino acids, between the sequence “FLLEP” and “WLCS” of SEQ ID NO:6. Such additional sequence can be identical to, substantially similar to, or different from, the sequence GEAGGDRP (portion of SEQ ID NO:4) or the sequence GEAGGDHP (portion of SEQ ID NO:8).

[0075] Likewise, it is contemplated that a naturally occurring human NR3B polypeptide can contain several additional amino acids (e.g. 1-20 additional amino acids, such as 7 additional amino acids), or no amino acids, between the sequence “WLCS” and “ELQEL” of SEQ ID NO:6. Such additional sequence can be identical to, substantially similar to, or different from, the sequence NGPGLQA (portion of SEQ ID NO:4) or the sequence NGPGVQA (portion of SEQ ID NO:8). Furthermore, it is contemplated that a naturally occurring human NR3B polypeptide does not contain the residues in SEQ ID NO:6 that extend beyond the corresponding residues from the C-terminus of SEQ ID NO:4 and 8, such as the sequence PPHSGRPGSQE (portion of SEQ ID NO:6).

[0076] A human NR3B polypeptide can contain C-terminal amino acid sequences that are not present in a sequence submitted to GenBank and annotated as a hypothetical protein most similar to rat ionotropic gluatmate receptor (L34938) with an ill-defined C-terminus (GenBank entry AC004528 and AAC12680; SEQ ID NOS:9 and 10). In particular, a human NR3B polypeptide can contain any or all of the C-terminal portion of SEQ ID NO:6 not also present in SEQ ID NO:10, such as the sequence XXXXXXXXXXXXXWKRARRAVDKERRVRFLLEPXXXXXXXXWLCSXXXXXXXELQEL ERRIEVARERLRQALVRRGQLLAQLGDSARHRPRRLLQARAAPAEAPPHSGRPGSQE where X can be any amino acid.

[0077] SEQ ID NOS:4 and 2 represent the predicted amino acid sequence of a naturally occurring rat NR3B polypeptide. SEQ ID NO:8 represents the predicted amino acid sequence of a naturally occurring mouse NR3B polypeptide.

[0078] An isolated nucleic acid molecule encoding SEQ ID NO:6 can have the nucleotide sequence designated SEQ ID NO:5, which represents a naturally occurring human NR3B cDNA sequence. The skilled person understands, however, that due to the degeneracy of the genetic code, SEQ ID NO:6 can also be encoded by a nucleotide sequence that differs from SEQ ID NO:5 at one or more codons.

[0079] Likewise, isolated nucleic acid molecules encoding SEQ ID NOS:4, 2 or 8 can have the nucleotide sequences designated SEQ ID NOS:3, 1 or 7, or degenerate variants thereof.

[0080] As shown in the alignment in FIG. 8, SEQ ID NOS:2, 4, 6 and 8 are highly homologous over their entire lengths. Because of this high degree of identity of NR3B polypeptides across these three mammalian species, it is expected that other naturally occurring mammalian NR3B polypeptides, such as NR3B polypeptides from non-human primates, mouse, rat, rabbit, bovine, porcine, ovine, canine or feline species, as well as naturally occurring NR3B polypeptides from other vertebrates, including fish, birds, reptiles and amphibians (e.g. Xenopus) will also exhibit a high degree of identity across their lengths with SEQ ID NO:6.

[0081] Using knowledge of the human, rat or mouse NR3B-encoding nucleic acid sequences and polypeptides disclosed herein, those skilled in the art can readily clone NR3B-encoding nucleic acids from other mammalian or vertebrate species using conventional cDNA or expression library screening methods, or using the polymerase chain reaction (PCR). Additionally, using knowledge of the human, rat or mouse NR3B-encoding nucleic acid sequences and polypeptides disclosed herein, those skilled in the art can readily determine cDNA and coding sequences form other species from an analysis of ESTs and genomic sequences present in available databases.

[0082] In contrast, SEQ ID NO:4 exhibits about 47% identity to rat NR3A, with much lower identity to other NMDA receptor subunits (i.e. 19.7% identity to rat NR1; 18.6% identity to rat NR2A). Therefore, the skilled person can readily distinguish an NR3B polypeptide from related receptor subunits based on sequence similarity.

[0083] For certain applications, an isolated nucleic acid molecule encoding an NR3B polypeptide need not encode the naturally occurring signal peptide sequence, which is cleaved in the mature polypeptide. The predicted signal peptide sequences of rat (SEQ ID NOS:2 and 4), human (SEQ ID NO:6) and mouse (SEQ ID NO:8) NR3B polypeptides are shown by overlining and the designation “SP” in FIG. 8. Accordingly, in one embodiment, an isolated nucleic acid molecule can encode an NR3B polypeptide in which some or all or of amino acids 1-51 or 1-53 of SEQ ID NOS:2, 4, 6 or 8 are not present. The skilled person can readily determine the boundaries of the signal peptide sequence from NR3B polypeptides and, if desired, replace these residues with another signal or sorting sequence.

[0084] An isolated nucleic acid molecule encoding an NR3B polypeptide can also be a splice variant form that differs from another form by one or more exons, thereby encoding a NR3B polypeptide that differs from another NR3B polypeptide by an insertion or deletion of one or more residues at one or more places in the polypeptide. NR3B splice variants can be expressed in a tissue or developmental stage-specific manner. Rat NR3B A2 (SEQ ID NO:2) and rat NR3B B4 (SEQ ID NO:4) are examples of splice variant forms that differ by containing (SEQ ID NO:4) or not containing (SEQ ID NO:2) the sequence VSVLRREVRTALGAP (portion of SEQ ID NO:4). An exemplary splice variant of a human NR3B differs from SEQ ID NO:6 by not containing the sequence LSLLRREARAPLGAP (portion of SEQ ID NO:6) or by not containing the sequence LSLLRREARAPLGAPN (portion of SEQ ID NO:6). An exemplary splice variant of a mouse NR3B differs from SEQ ID NO:8 by not containing the sequence LSVLRREVRAPLGAR (portion of SEQ ID NO:8) or by not containing the sequence LSVLRREVRAPLGARR (portion of SEQ ID NO:8). The skilled person can readily determine additional splice variants of these and other NR3B polypeptides.

[0085] An isolated nucleic acid molecule encoding an NR3B polypeptide can also have one or more minor modifications to the naturally occurring sequence, such as one or more substitutions additions or deletions. Such modifications can be advantageous, for example, in enhancing the stability, bioavailability, bioactivity or immunogenicity of the polypeptide, or to facilitate its purification.

[0086] Substitutions to an NR3B amino acid sequence can either be conservative or non-conservative. Conservative amino acid substitutions include, but are not limited to, substitution of an apolar amino acid with another apolar amino acid (such as replacement of leucine with an isoleucine, valine, alanine, proline, tryptophan, phenylalanine or methionine); substitution of a charged amino acid with a similarly charged amino acid (such as replacement of a glutamic acid with an aspartic acid, or replacement of an arginine with a lysine or histidine); substitution of an uncharged polar amino acid with another uncharged polar amino acid (such as replacement of a serine with a glycine, threonine, tyrosine, cysteine, asparagine or glutamine); or substitution of a residue with a different functional group with a residue of similar size and shape (such as replacement of a serine with an alanine; an arginine with a methionine; or a tyrosine with a phenylalanine).

[0087] Additions to an NR3B amino acid sequence include, but are not limited to, the addition of “tag” sequences, which are conveniently added at the N- or C-termini, after the signal peptide, or within extracellular or intracellular loops. Such tag sequence include, for example, epitope tags, histidine tags, glutathione-S-transferase (GST), fluorescent proteins (e.g. Enhanced Green Fluorescent Protein (EGFP)) and the like. Such additional sequences can be used, for example, to facilitate expression, purification or characterization of an NR3B polypeptide.

[0088] Deletions to an NR3B amino acid sequence include, but are not limited to, deletion of signal peptide residues, and deletion of residues at the N- or C- termini that are not critical for function. Deleted sequences can optionally be replaced by tag sequences or fusion sequences, as described previously.

[0089] Modifications to an encoded NR3B amino acid sequence, such as modifications to any of SEQ ID NOS:2, 4, 6 or 8, can be randomly generated, such as by random insertions, deletions or substitutions of nucleotides in a nucleic acid molecule encoding the polypeptide. Alternatively, modifications can be directed, such as by site-directed mutagenesis of a nucleic acid molecule encoding the polypeptide.

[0090] Guidance in modifying the sequence of an NR3B polypeptide while retaining biological activity can be provided by the alignment of the sequence of the NR3B orthologs from human, rat and mouse shown in FIG. 8. It is well known in the art that evolutionarily conserved amino acid residues are more likely to be important for maintaining biological activity than less well-conserved residues. Thus, it would be expected that substituting a residue that is highly conserved among NR3B polypeptides across species with a non-conserved residue may be deleterious, whereas making the same substitution at a residue which varies among species would likely not have a significant effect on biological activity.

[0091] Additionally, guidance in modifying amino acid residues of an NR3B polypeptide while retaining a desired biological activity can be provided by structure-function studies of known NMDA receptor subunits, which share an overall transmembrane topology and domain structure with NR3B. By analogy to other subunits, the ligand binding domain of NR3B is predicted to be formed by the extracellular S1 domain before the first membrane spanning region (M1) and by the extracellular S2 domain between membrane spanning regions M3 and M4 (FIG. 8). The second membrane domain (M2, or P-loop) is predicted to line the ion channel pore (see FIG. 8). Meddows et al., J. Biol. Chem. 276:18795-18803 (2001) have also determined that retention of the N-terminal residues of the NMDA receptor subunit NR1 (i.e. amino acids 1-380 of NR1) is important for subunit oligomerization, whereas the M4 domain and C-terminal residues (i.e. amino acids 811-938 of NR1) are dispensible for oligomerization but required for functional channel formation. The skilled person could apply this knowledge to predict the effect of various modifications within the above-described structural and functional domains on NR3B biological activity.

[0092] Computer programs well known in the art can also provide guidance in predicting which amino acid residues can be modified without abolishing a topological or functional feature of an NR3B polypeptide.

[0093] The invention also provides an isolated nucleic acid molecule that encodes a functional fragment of an NR3B polypeptide. As used herein, the term “functional fragment” refers to a portion of a full-length NR3B polypeptide that retains at least one biological activity characteristic of the full-length polypeptide. A functional fragment can contain, for example, at least about 6, 8, 10, 12, 15, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 110, 125, 150, 200, 250, 300, 400, 500, 600, 700, 800, 900, 950 or more amino acids of an NR3B polypeptide.

[0094] For example, a functional fragment of an NR3B polypeptide can retain the ability to bind glycine. As shown in FIGS. 1 and 9, the residues of the S1 and S2 regions considered to be important in binding glycine are known. Thus, a functional fragment can contain all or part of the S1 and/or S2 domains of rat, human or mouse NR3B (see FIG. 8), and optionally further contain the naturally occurring NR3B intervening sequence, such as membrane regions M1-M3.

[0095] An exemplary NR3B functional fragment that binds glycine can contain SEQ ID NO:27 and/or SEQ ID NO:35 and/or SEQ ID NO:43. Advantageously, a chimeric polypeptide containing all or a portion of a different NMDA receptor subunit (e.g. NR1, NR2A-D or NR3A), with the glycine binding domain (e.g. the S1 and/or S2 regions; or SEQ ID NO:27 and/or SEQ ID NO:35 and/or SEQ ID NO:43) of NR3B replacing the corresponding region of the NMDA receptor subunit, can be constructed. Likewise, a chimeric polypeptide containing all or a portion of an NMDA receptor subunit (e.g. NR1, NR2A-D or NR3B), with the glycine binding domain (e.g. the S1 and/or S2 regions; or SEQ ID NO:26 and/or SEQ ID NO:34 and/or SEQ ID NO:42) of NR3A replacing the corresponding region of the NMDA receptor subunit, can be constructed. Such a functional fragment, or a chimeric polypeptide containing such a fragment, can be used, for example, in screening applications described further below to detect excitatory glycine receptor ligands, agonists and antagonists. Additionally, such a functional fragment can be used in therapeutic applications in which it is desirable to compete with an endogenous receptor for binding to agonist. Methods for making and testing chimeric glutamate receptor polypeptides are described, for example, in Villmann et al., Eur. J. Neurosci. 11:1765-1778 (1999).

[0096] A functional fragment of NR3B can also retain the ability to oligomerize with other NMDA receptor subunits, such as NR1, and optionally NR2. By analogy to NR1, such a fragment can retain all or most of the N-terminal region before the S1 domain, which is predicted to be important for oligomerization (see Meddows et al., supra (2001)).

[0097] A further exemplary functional fragment of NR3B can retain the ability to insert into the membrane or form a channel pore by retaining some or all of the membrane regions (M1-M4). Such fragments can be used, for example, to compete with or disrupt the structure of the naturally occurring NR3B.

[0098] Another exemplary functional fragment of NR3B can retain the ability to interact with intracellular proteins, such as effector proteins, by retaining some or all of the intracellular region C-terminal to M4. Such fragments can be used, for example, in binding assays to identify polypeptides that interact with NR3B, which can then themselves be used as targets in screening assays; and also can be used to compete with naturally occurring NR3B for binding to effector polypeptides.

[0099] Accordingly, the invention provides an isolated nucleic acid molecule that encodes an NR3B functional fragment that contains the extracellular domain of an NR3B polyepeptide N-terminal to the S1 domain (with or without the signal peptide), and/or the S1 domain, and/or the M1 domain, and/or the M2 domain, and/or the and/or the M3 domain, and/or the S2 domain, and/or the M4 domain, and/or the intracellular domain C-terminal to the M4 domain. The boundaries of these domains for several mammalian NR3B polypeptides are shown in FIG. 8. The skilled person can determine appropriate functional fragments of an NR3B polypeptide for use in a particular application.

[0100] The biological activities of NR3B polypeptides and functional fragments can be determined or confirmed by methods known in the art and described further in the Examples. For example, the ability of an NR3B polypeptide or functional fragment to act as a subunit of an excitatory glycine receptor can be tested by recombinantly expressing an NR3B polypeptide in an appropriate cell (e.g. a Xenopus oocyte or mammalian cell) in the presence of a suitable amount of another endogenous or exogenous NMDAR subunit (e.g. an NR1 subunit, an NR2 subunit, or an NR3A subunit, or any combination thereof that includes an NR1 subunit), and detecting currents evoked in a dose-dependent fashion by addition of glycine. Other suitable methods for detecting the ability of an NR3B polypeptide to act as a subunit of an excitatory glycine receptor are known in the art and described further below with respect to screening assays.

[0101] The ability of an NR3B polypeptide or functional fragment to oligomerize with an NR1 and/or an NR2 and/or an NR3A polypeptide can be assayed, for example, by a functional assay to measure excitatory ionic responses of an NR3B/NR1 receptor to glycine, as described above, or alternatively by co-expressing the polypeptides and detecting NR3B/NR1 polypeptide association. Various assays for detecting polypeptide associations are well known in the art and include, for example, co-immunoprecipitation assays (see, for example, Meddows et al., supra (2001)), two-hybrid assays, GST pull-down assays, protein chip proteomic array analysis (e.g. ProteinChip™ System from Ciphergen Biosystems, which can be used in tandem with mass spectrometry analysis for sequence or structure determination) and the like, using an NR3B polypeptide or functional fragment.

[0102] The ability of an NR3B polypeptide or functional fragment to bind glycine can be also detected by a functional assay to measure excitatory ionic responses of an NR3B/NR1 receptor to glycine, as described above, or alternatively by a ligand binding assay. Various direct and competitive ligand binding assays, such as those described above with respect to oligomerization, and assays described below with respect to screening, are well known in the art and can be used to determine the ability of an NR3B polypeptide or functional fragment to bind glycine.

[0103] Further provided are isolated polynucleotides containing at least 17 contiguous nucleotides of an invention NR3B nucleic acid molecule or of its complement. An isolated polynucleotide can thus contain at least 18, 19, 20, 22, or at least 25 contiguous nucleotides, such as at least 30, 40, 50, 60, 70, 80, 90, 100, 125, 150, 175, 200, 225, 250, 275, 300, 350, 400, 500, 600, 700, 800, 1000, 1500, 2000, 2500, 3000, 3500 or more contiguous nucleotides from the reference nucleotide sequence, up to the full length sequence. An invention polynucleotide can be single or double stranded, represent the sense or antisense strand, and contain either coding or non-coding sequence or both. An invention polynucleotide can, but need not, encode a biologically active polypeptide and can, but need not, be inserted into a vector.

[0104] In one embodiment, the isolated polynucleotide comprises at least 17 contiguous nucleotides of any of SEQ ID NOS:1, 3, 5 or 7 or the complement thereof. Such polynucleotides are of sufficient length and complexity to be able to specifically hybridize to an NR3B-encoding nucleic acid molecule under highly stringent hybridization conditions. Therefore, the invention polynucleotides can advantageously be used, for example, as probes to detect the presence, abundance or fidelity of NR3B-encoding nucleic acid molecules in a sample; as NR3B-specific sequencing or PCR primers; as antisense, RNA interference or ribozyme reagents for use in ex vivo or in vivo gene therapy applications to block expression of NR3B in a cell, as described in more detail below; or in other applications known to those skilled in the art in which hybridization to an NR3B-encoding nucleic acid molecule is desirable. In certain applications, polynucleotides that distinguish a splice variant form of an NR3B receptor are useful, such as polynucleotides containing the region of the rat NR3B B4 form not present in the NR3B A2 form.

[0105] Specific hybridization refers to the ability of a nucleic acid molecule to hybridize to the reference nucleic acid molecule without hybridization under the same conditions with nucleic acid molecules that are not the reference molecule, such as a nucleic acid molecule encoding another NMDA receptor subunit. Moderately stringent hybridization conditions are conditions equivalent to hybridization of filter-bound nucleic acid in 50% formamide, 5×Denhart's solution, 5×SSPE, 0.2% SDS at 42° C., followed by washing in 0.2×SSPE, 0.2% SDS, at 50°. Highly stringent conditions are conditions equivalent to hybridization of filter-bound nucleic acid in 50% formamide, 5×Denhart's solution, 5×SSPE, 0.2% SDS at 42° C., followed by washing in 0.2×SSPE, 0.2% SDS, at 65° C. Other suitable moderately stringent and highly stringent hybridization buffers and conditions are well known to those of skill in the art and are described, for example, in Sambrook et al., Molecular Cloning: A Laboratory Manual, 3rd ed., Cold Spring Harbor Press, Plainview, N.Y. (2001) and in Ausubel et al. (Current Protocols in Molecular Biology (Supplement 47), John Wiley & Sons, New York (1999)).

[0106] In one embodiment, the invention provides a primer pair containing two isolated polynucleotides as set forth above. The primer pair can be used, for example, to amplify an NR3B-encoding nucleic acid molecule by the polymerase chain reaction (PCR). A suitable primer pair can contain an isolated polynucleotide containing at least 17 contiguous nucleotides of the sense strand of an invention NR3B nucleic acid molecule, and an isolated polynucleotide containing at least 17 contiguous nucleotides of the antisense strand of an invention NR3B nucleic acid molecule. The skilled person can determine an appropriate primer length and sequence composition for the intended application.

[0107] NR3B nucleic acid molecules (including nucleic acid molecules encoding NR3B polypeptides, functional fragments thereof and polynucleotides, as described above) can optionally contain exogenous nucleotide sequences including, for example, sequences that facilitate identification or purification of the molecule, and sequences that facilitate cloning, such as restriction endonuclease recognition sites.

[0108] NR3B nucleic acid molecules can be produced or isolated by methods known in the art. The method chosen will depend on the type of nucleic acid molecule one intends to isolate. Those skilled in the art, based on knowledge of the nucleotide sequences disclosed herein, can readily isolate NR3B nucleic acid molecules as genomic DNA, as full-length cDNA or desired fragments therefrom, or as full-length mRNA or cRNA or desired fragments therefrom, by methods known in the art.

[0109] It will be appreciated that an invention NR3B polypeptide, functional fragment or peptide does not consist of the exact sequence of an amino acid sequence set forth in a publically available database, or of the exact amino acid sequence of a translated product of a nucleic acid molecule set forth in a publically available database. Likewise, an invention nucleic acid molecule encoding a NR3B polypeptide or functional fragment, or an NR3B polynucleotide, does not consist of the exact sequence of a nucleotide sequence set forth in publically available databases, including but not limited to Expressed Sequence Tags (ESTs), Sequence Tagged Sites (STSs) and genomic fragments deposited in public databases such as the GenBank nr, dbest, dbsts and gss databases.

[0110] Specifically excluded from the invention polypeptides and nucleic acid molecules are molecules having the exact sequence of any of the following: the human EST sequence designated SEQ ID NO:13 (GenBank Accession No. AL359933); fragments of human chromosome 19 genomic sequences (e.g. GenBank Accession No. AC004528), such as the predicted cDNA sequence designated SEQ ID NO:9 which encodes a protein designated as a hypothetical human protein most similar to rat ionotropic gluatmate receptor, and the encoded polypeptide, SEQ ID NO:10 (GenBank Accession No. AAC12680); the mouse EST sequence designated SEQ ID NO:11 (GenBank Accession No. BC005494) and its predicted encoded polypeptide designated SEQ ID NO:12 (GenBank Accession No. AAH05494.1); and deposited fragments of mouse chromosome 10 genomic sequences (e.g. GenBank Accession No. AC087114).

[0111] Since one of skill in the art will realize that the above-recited excluded sequences may be revised in the database at a later date, it is intended that the above-recited sequences are excluded as they stand on the priority date of this application.

[0112] Isolated NR3B nucleic acid molecules can be prepared or isolated by methods well known in the art. The method chosen will depend on factors such as the type and size of the nucleic acid molecule; whether or not it encodes a biologically active polypeptide; and the source of the nucleic acid molecule. Such methods are described, for example, in Sambrook et al., supra (2001) and in Ausubel et al., supra (1999).

[0113] One useful method for producing an isolated NR3B nucleic acid molecule involves amplification of the nucleic acid molecule using the polymerase chain reaction (PCR) and specific primers and, optionally, purification of the resulting product by gel electrophoresis. Either PCR or reverse-transcription PCR (RT-PCR) can be used to produce a nucleic acid molecule having any desired nucleotide boundaries. Desired modifications to the nucleic acid sequence can also be conveniently introduced by choosing an appropriate primer with one or more additions, deletions or substitutions. Such nucleic acid molecules can be amplified exponentially starting from as little as a single gene or mRNA copy, from any cell, tissue or species of interest.

[0114] An isolated NR3B nucleic acid molecule can also be prepared by screening a library, such as a genomic library, cDNA library or expression library, with a detectable NR3B nucleic acid molecule or antibody. Human libraries, and libraries from a large variety of other species, are commercially available or can be produced from species or cells of interest. The library clones identified as containing NR3B nucleic acid molecules can be isolated, subcloned or sequenced by routine methods. From an initially identified fragment, nucleic acid molecules encoding full-length polypeptides can be obtained, if desired, by a variety of methods well-known in the art, such as 5′ or 3′ RACE.

[0115] Furthermore, an isolated NR3B nucleic acid molecule can be produced by synthetic means. For example, a single strand of a nucleic acid molecule can be chemically synthesized in one piece, or in several pieces, by automated synthesis methods known in the art. The complementary strand can likewise be synthesized in one or more pieces, and a double-stranded molecule made by annealing the complementary strands. Direct synthesis is particularly advantageous for producing relatively short molecules, such as probes and primers, and nucleic acid molecules containing modified nucleotides or linkages.

[0116] The invention also provides a vector containing an isolated NR3B nucleic acid molecule. The vectors of the invention are useful, for example, for subcloning and amplifying NR3B nucleic acid molecules, and for recombinantly expressing NR3B polypeptides and functional fragments thereof. A vector of the invention can include a variety of elements useful for cloning and/or expression of the encoded nucleic acid molecule in the desired host cell, such as promoter and/or enhancer sequences, which can provide for constitutive, inducible or cell-specific RNA transcription; transcription termination and RNA processing signals, including polyadenylation signals, which provide for stability of a transcribed mRNA sequence; an origin of replication, which allows for proper episomal replication; selectable marker genes, such as a neomycin or hygromycin resistance gene, useful for selecting stable or transient transfectants in mammalian cells, or an ampicillin resistance gene, useful for selecting transformants in prokaryotic cells; and versatile multiple cloning sites for inserting nucleic acid molecules of interest.

[0117] Cloning vectors of the invention include, for example, viral vectors such as a bacteriophage, a baculovirus or a retrovirus; cosmids or plasmids; and, particularly for cloning large nucleic acid molecules, bacterial artificial chromosome vectors (BACs) and yeast artificial chromosome vectors (YACs). Such vectors are commercially available, and their uses are well known in the art.

[0118] If it is desired to express NR3B RNA transcripts or polypeptides, an invention nucleic acid molecule can be operatively linked to a promoter of RNA transcription. The term “operatively linked,” as used herein, is intended to mean that the nucleic acid molecule is positioned with respect to the endogenous promoter, or heterologous promoter, in such a manner that the promoter will direct the transcription of RNA using the nucleic acid molecule as a template. Methods for operatively linking a nucleic acid to a desired promoter are well known in the art and include, for example, cloning the nucleic acid into a vector containing the desired promoter, or appending the promoter to a nucleic acid sequence using PCR.

[0119] Thus, an invention nucleic acid molecule operatively linked to a promoter of RNA transcription can be used to express NR3B transcripts and polypeptides in a desired host cell, or in an in vitro system, such as an extract or lysate that supports transcription and translation.

[0120] Contemplated promoters and expression vectors provide for expression in bacterial cells, yeast cells, insect cells, amphibian cells, mammalian cells (including human, non-human primate and rodent cells) and other vertebrate cells. A variety of promoters and expression vectors suitable for such purposes are commercially available, and can be further modified, if desired, to include appropriate regulatory elements to provide for the desired level of expression or replication in the host cell.

[0121] For use in the gene therapy applications described further below, an invention nucleic acid molecule can be incorporated into suitable gene therapy vector, such as a viral vector or plasmid. Viral based vectors are advantageous in being able to introduce relatively high levels of a heterologous nucleic acid into a variety of cells, including nondividing cells.

[0122] Suitable viral vectors for gene therapy applications are well known in the art, and include, for example, Herpes simplex virus vectors (U.S. Pat. No. 5,501,979), Vaccinia virus vectors (U.S. Pat. No. 5,506,138), Cytomegalovirus vectors (U.S. Pat. No. 5,561,063), Modified Moloney murine leukemia virus vectors (U.S. Pat. No. 5,693,508), adenovirus vectors (U.S. Pat. Nos. 5,700,470 and 5,731,172), adeno-associated virus vectors (U.S. Pat. No. 5,604,090), constitutive and regulatable retrovirus vectors (U.S. Pat. Nos. 4,405,712; 4,650,764 and 5,739,018, 5,646,013, 5,624,820, 5,693,508 and 5,674,703), papilloma virus vectors (U.S. Pat. Nos. 5,674,703 and 5,719,054), lentiviral vectors (Kafri et al., Mol. Ther. 1:516-521 (2000), and the like. For targeting neural cells in the treatment of neuronal diseases, adenoviral vectors, Herpes simplex virus vectors and lentiviral vectors are particularly useful.

[0123] The invention also provides a cell containing an isolated NR3B nucleic acid molecule. Such a cell need not express a recombinant NR3B polypeptide or fragment for use in cloning procedures. However, a cell can optionally express an NR3B polypeptide or functional fragment encoded by the nucleic acid molecule. Such cells can be used in a variety of applications including, for example, screening for agonists, antagonists and ligands of excitatory glycine receptors, as described further below; as a source to isolate recombinantly expressed NR3B polypeptides; for identifying additional cellular molecules, such as additional receptor subunits or intracellular proteins that associate with NR3B; and in other applications known to those skilled in the art.

[0124] Optionally, a cell that recombinantly expresses an NR3B polypeptide can further endogenously or recombinantly express at least one other NMDA subunit, such as an NR1 subunit. As disclosed herein, co-expression of an NR3B and an NR1 polypeptide results in the formation of excitatory glycine receptors. NR1 polypeptides and encoding nucleic acid molecules from various species are known in the art, and include the naturally occurring NR1 polypeptides from human, rat, mouse, duck, fish and Xenopus having the nucleotide and predicted amino acid sequences set forth in Table 1: TABLE 1 SUBUNIT ACCESSION NUMBER Human NR1 D13515 Human NR1-1 L13266 Human NR1-2 L13267 Human NR1-3 L13268 Human NR1-3b AF015730 Human NR1-4b AF015731 Rat NR1 U11418 Rat NR1 X63255 Rat NR1-2b U08264 Rat NR1-3a U08265 Rat NR1-3b U08266 Rat NR1-4a U08267 Rat NR1-4b U08268 Mouse NR1 D10028 Duck NR1 D83352 Fish NR1 AF060557 Rat NR1-1a U08261 Rat NR1-2a U08262 Rat NR1-1b U08263 Xenopus NR1 X94081

[0125] The skilled person will appreciate that for use in the methods described herein, a co-expressed NR1 polypeptide can have modifications to the naturally occurring sequence, or be a functional fragment of the naturally occurring sequence, so long as the desired NR1 biological activity is retained. An exemplary modification of a naturally occurring NR1 polypeptide that does not affect biological activity is the addition of an epitope tag to facilitate identification in procedures such as immunolocalization and immunoprecipitation. NR1 biological activities include, for example, the ability to oligomerize with other NMDA subunits, including NR2A-D, NR3B and NR3A; the ability to bind glycine; the ability to form excitatory glycine receptors in association with either NR3B or NR3A; and the ability to form NMDA- and glycine-responsive receptors in association with NR2 subunits. As described above with respect to NR3B polypeptides and functional fragments, the skilled person can readily make NR1 molecules with sequences that differ from the naturally occurring sequence and test such molecules to confirm that a desired NR1 biological activity is retained.

[0126] Exemplary host cells that can be used to recombinantly express receptor polypeptides and fragments include mammalian primary cells; established mammalian cell lines, such as COS, CHO, HeLa, NIH3T3, HEK 293-T and PC12 cells; amphibian cells, such as Xenopus embryos and oocytes; and other vertebrate cells. Exemplary host cells also include insect cells (e.g. Drosophila), yeast cells (e.g. S. cerevisiae, S. pombe, or Pichia pastoris) and prokaryotic cells (e.g. E. coli).

[0127] Also provided are extracts of recombinant cells that express NR3B at the cell membrane, wherein the extract contains the cell membrane. Advantageously, NR3B is expected to retain a biologically active conformation in a membrane extract, but is partially purified away from other cellular components that may be undesirable for certain applications. Cell membrane extracts can be prepared by methods known in the art (see, for example Das et al., Nature 393:377-381 (1998)) and can be used in many of the screening assays described herein.

[0128] Methods for introducing a recombinant nucleic acid molecule into a host cell are well known in the art. The choice of method will depend on the host cell, the type of nucleic acid molecule, and the intended application of the host cell. Suitable methods include, for example, various methods of transfection such as calcium phosphate, DEAE-dextran and lipofection methods; viral transduction; electroporation; and microinjection.

[0129] Animal model systems can be useful for elucidating normal and pathological functions of NR3B polypeptides, and for determining the efficacy and safety of potential therapeutic compounds that target excitatory glycine receptors. Accordingly, the invention also provides a transgenic non-human animal that contains an NR3B nucleic acid molecule. Such transgenic animals can express a nucleic acid molecule encoding an invention NR3B polypeptide or functional fragment, including a functional fragment that competes with or inhibits the function of the naturally occurring NR3B, as described previously. Alternatively, transgenic animals can express an invention NR3B polynucleotide that prevents effective translation of the naturally occurring NR3B polypeptide, such as an antisense, ribozyme, RNAi or similar construct. By employing suitable inducible and/or tissue specific regulatory elements, NR3B expression or activity in the transgenic animal can be restricted to specific cell types, developmental stages, or induction conditions.

[0130] Any of a variety of methods known in the art can be used to introduce a desired transgene into animals to produce the founder lines of transgenic animals (see, for example, Hogan et al., Manipulating the Mouse Embryo: A Laboratory Manual, second ed., Cold Spring Harbor Laboratory (1994), U.S. Pat. Nos. 5,602,299; 5,175,384; 6,066,778; and 6,037,521). Such techniques include, but are not limited to, pronuclear microinjection (U.S. Pat. No. 4,873,191); retrovirus mediated gene transfer into germ lines (Van der Putten et al., Proc. Natl. Acad. Sci. USA 82:6148-6152 (1985)); gene targeting in embryonic stem cells (Thompson et al., Cell 56:313-321 (1989)); electroporation of embryos (Lo, Mol Cell. Biol. 3:1803-1814 (1983)); and sperm-mediated gene transfer (Lavitrano et al., Cell 57:717-723 (1989)). Once the founder animals are produced, they can be bred to produce colonies of the particular animal by methods known in the art.

[0131] In another embodiment, the invention provides NR3B-deficient non-human animals, or NR3B “knock-out” animals. Methods of deleting all or a portion of a gene so as to alter or prevent expression of the naturally occurring polypeptide are well known in the art. Gene knockout by homologous recombination is described, for example, in Capecchi et al., Science 244:1288 (1989), and in U.S. Pat. Nos. 5,616,491, 5,750,826, and 5,981,830. Methods of making and using an NR3A knockout mouse are described in Das et al., Nature 393:377-381 (1998). Analogous targeting vectors and methods are expected to be useful in generating NR3B knockout animals. FIG. 10 describes a targeting construct suitable for use in generating an NR3B knockout mouse.

[0132] The invention also provides a method for detecting a nucleic acid molecule encoding an NR3B polypeptide in a sample. Because of the critical role NMDA receptors play in neurologic disorders, the method can be used, for example, to diagnose or prognose a pathological condition mediated, in part, by altered expression, abundance or integrity of an NR3B nucleic acid molecule. Such conditions include, for example, acute neurologic conditions and chronic neurodegenerative diseases described further below. The detection method can also be used to identify additional regions of the nervous system in which NR3B is normally or pathologically expressed. Such information can be valuable in determining additional diagnostic and therapeutic applications for the invention molecules and methods described herein. Furthermore, the detection method can also be used to identify additional naturally occurring splice variants of NR3B receptors, and NR3B-encoding nucleic acid molecules from other species of interest, such as veterinary and laboratory animals.

[0133] In one embodiment, the detection method is practiced by contacting a sample with one or more NR3B polynucleotides and detecting specific hybridization to the polynucleotide. Suitable hybridization conditions for detecting specific hybridization will depend on the detection format, and are well known in the art. Exemplary conditions useful for filter-based assays have been described previously. Example II provides exemplary conditions suitable for in situ hybridization assays. Suitable conditions for PCR-based detection methods are also well known in the art and described, for example, in Sambrook et al., supra (2001) and in Ausubel et al., supra (1999).

[0134] As used herein, the term “sample” is intended to mean any biological fluid, cell, tissue, organ or portion thereof that contains or potentially contains an NR3B nucleic acid molecule or polypeptide. For example, a sample can be a histologic section of a specimen obtained by biopsy, or cells that are placed in or adapted to tissue culture. A sample further can be a subcellular fraction or extract, or a crude or substantially pure nucleic acid or protein preparation. A sample can be prepared by methods known in the art suitable for the particular format of the detection method employed.

[0135] The methods of detecting an NR3B nucleic acid molecule in a sample can be either qualitative or quantitative, and can detect the presence, abundance, integrity or structure of the nucleic acid molecule, as desired for a particular application. Suitable hybridization-based assay methods include, for example, in situ hybridization, which can be used to detect altered chromosomal location of the nucleic acid molecule, altered gene copy number, and RNA abundance, depending on the assay format used. Other hybridization methods include, for example, Northern blots and RNase protection assays, which can be used to determine the abundance and integrity of different RNA splice variants, and Southern blots, which can be used to determine the copy number and integrity of DNA. A hybridization probe can be labeled with any suitable detectable moiety, such as a radioisotope, fluorochrome, chemiluminescent marker, biotin, or other detectable moiety known in the art that is detectable by analytical methods.

[0136] Suitable amplification-based detection methods are also well known in the art, and include, for example, qualitative or quantitative polymerase chain reaction (PCR); reverse-transcription PCR (RT-PCR); single strand conformational polymorphism (SSCP) analysis, which can readily identify a single point mutation in DNA based on differences in the secondary structure of single-strand DNA that produce an altered electrophoretic mobility upon non-denaturing gel electrophoresis; and coupled PCR, transcription and translation assays, such as a protein truncation test, in which a mutation in DNA is determined by an altered protein product on an electrophoresis gel. The amplified nucleic acid molecule can be sequenced to detect mutations and mutational hot-spots, and specific PCR-based assays for large-scale screening of samples to identify such mutations can be developed.

[0137] The invention also provides isolated NR3B polypeptides and functional fragments therefrom, having amino acid sequences as described above with respect to polypeptides encoded by invention nucleic acid molecules. NR3B polypeptides and functional fragments can be used, for example, in therapeutic applications in which such polypeptides and fragments are administered onto or into cells; in screening assays to identify ligands, agonists and antagonists of excitatory glycine receptors; in research applications to identify additional NR3B-associating polypeptides; to raise antibodies for use in diagnostic and prognostic methods; to affinity purify antibodies and ligands; and in other applications known to those skilled in the art.

[0138] An isolated NR3B polypeptide of the invention can optionally contain amino acids with various chemical or enzymatic modifications with respect to naturally occurring amino acids. Such modifications can enhance the stability, bioactivity, immunogenicity or other advantageous property of an invention polypeptide. Thus, a polypeptide can contain an amino acid modified by replacement of hydrogen by an alkyl, acyl, or amino group; by esterification of a carboxyl group with a suitable alkyl or aryl moiety; by alkylation of a hydroxyl group to form an ether derivative; by phosphorylation or dephosphorylation of a serine, threonine or tyrosine residue; by N- or O-linked glycosylation; by iodination; by radiolabeling; or the like. A polypeptide can also include a modified amino acids such as hydroxyproline or carboxyglutamate, or a D-amino acid in place of its corresponding L-amino acid. Those skilled in the art can determine an appropriate amino acid modification for a given application.

[0139] In another embodiment, the invention provides an isolated NR3B peptide. An exemplary NR3B peptide contains at least 8 contiguous amino acids of a naturally occurring NR3B polypeptide, such as at least 8 contiguous amino acids of SEQ ID NOS:2, 4, 6 or 8. Such a peptide can contain, for example, at least about 10, 12, 15, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 110, 125, 150, 200, 250, 300, 400, 500, 600, 700, 800, 900, 950 or more amino acids, up to the full length of the reference polypeptide. A peptide of at least about 8 amino acids can be used, for example, as an immunogen to raise antibodies specific for an NR3B polypeptide, or as an antigen to purify antibodies specific for an NR3B polypeptide. When used as an antigen, an invention peptide can be attached to a carrier molecule such as bovine serum albumin (BSA) or keyhole limpet hemocyanin (KLH).

[0140] Peptides that are likely to be antigenic or immunogenic can be predicted using methods and algorithms known in the art and described, for example, by Irnaten et al., Protein Eng. 11:949-955 (1998), and Savoie et al., Pac. Symp. Biocomput. 1999:182-189 (1999). Immunogenicity of the peptides of the invention can be determined by methods known in the art, such as assay of a delayed-type hypersensitivity response in an animal sensitized to a NR3B polypeptide, or by elicitation of antibodies specific for NR3B polypeptides. Likewise, antigenicity of the peptides of the invention can be determined by methods known in the art, such as by ELISA analysis, as described, for example, in Harlow and Lane, Antibodies: A Laboratory Manual, Cold Spring Harbor Laboratory Press (1988).

[0141] The isolated NR3B polypeptides, functional fragments and peptides of the invention can be prepared by methods known in the art, including biochemical, recombinant and synthetic methods. For example, polypeptides can be purified by routine biochemical methods from cells or tissues that express the polypeptide. The detection methods disclosed herein can be adapted for determining which cells or tissues are appropriate starting materials. Biochemical purification can include, for example, steps such as solubilization of the appropriate cells, size or affinity chromatography, electrophoresis, and immunoaffinity procedures. The methods and conditions for biochemical purification of a polypeptide of the invention can be chosen by those skilled in the art, and purification monitored, for example, by an ELISA assay or a functional assay.

[0142] An NR3B polypeptide, functional fragment or peptide having any desired boundaries can also be produced by recombinant methods. Recombinant methods involve expressing a nucleic acid molecule encoding the desired polypeptide or fragment in a host cell or cell extract, and isolating the recombinant polypeptide or fragment, such as by routine biochemical purification methods described above. To facilitate identification and purification of the recombinant polypeptide, it is often desirable to insert or add, in-frame with the coding sequence, nucleic acid sequences that encode epitope tags, polyhistidine tags, glutathione-S-transferase (GST) domains, fluorescent proteins (e.g. EGFP) and the like. Methods for producing and expressing recombinant polypeptides in vitro and in prokaryotic and eukaryotic host cells are well known in the art.

[0143] Thus, the invention provides a method of producing an NR3B polypeptide or functional fragment either in vitro or in a cell, by expressing a nucleic acid molecule encoding the polypeptide or fragment under appropriate conditions. Optionally, the polypeptide or fragment so produced can be partially purified, such as obtained as a membrane extract.

[0144] The invention NR3B polypeptide fragments and peptides can also be produced, for example, by enzymatic or chemical cleavage of the full-length polypeptide. Methods for enzymatic and chemical cleavage and for purification of the resultant peptide fragments are well known in the art (see, for example, Deutscher, Methods in Enzymology, Vol. 182, “Guide to Protein Purification,” San Diego: Academic Press, Inc. (1990)).

[0145] Depending on the intended application, the isolated NR3B polypeptide or functional fragment can optionally be isolated in, or reconstituted into, a natural or artificial lipid bilayer, such as a cell membrane or liposome, with or without other cellular components. Thus, in one embodiment, the invention provides an isolated NR3B polypeptide, further comprising a membrane, and optionally further comprising an NMDA receptor subunit, such as an NR1 subunit. Membrane-associated NR3B polypeptides and functional fragments are useful in applications in which structural integrity of the subunit and/or excitatory glycine receptor are important, such as in the screening assays described herein.

[0146] The invention also provides an antibody or antigen binding fragment thereof which specifically binds an NR3B polypeptide. Such antibodies, which include polyclonal, monoclonal, chimeric, bifunctional, and humanized antibodies, can be used, for example, to affinity purify an NR3B polypeptide or functional fragment; to detect cellular polypeptides, including NMDA receptor subunits, that associate with NR3B; and in therapeutic, diagnostic and research applications known to those skilled in the art and described further below. An antibody that “specifically binds” and NR3B polypeptide binds with high affinity to a NR3B polypeptide in a binding assay, such as an immunoblot or ELISA assay, without substantial cross-reactivity with other polypeptides such as other NMDA receptor subunits.

[0147] An “antigen binding fragment” of an antibody of the invention includes, for example, individual heavy or light chains and fragments thereof, such as VL, VH and Fd; monovalent fragments, such as Fv, Fab, and Fab′; bivalent fragments such as F(ab′)₂; single chain Fv (scFv); and Fc fragments. Antigen binding fragments include, for example, fragments produced by protease digestion or reduction of an antibody, as well as fragments produced by recombinant DNA methods known to those skilled in the art.

[0148] In one embodiment, the invention provides antibodies and antigen binding fragments thereof that specifically bind an NR3B polypeptide containing an amino acid sequence designated SEQ ID NO:2, 4, 6 or 8. For certain applications, such as to antagonize receptor activity, antibodies that bind an extracellular portion of NR3B are desirable, including antibodies that bind the region N-terminal to the S1 domain, the S1 domain, the S2 domain, or the region between the M3 and M4 domains. For other applications, antibodies that distinguish a splice variant form of an NR3B receptor are useful, such as antibodies that bind the 15 amino acid portion of rat NR3B B4 not present in the NR3B A2 form.

[0149] The antibodies of the invention can be produced by any suitable method known in the art. For example, a NR3B polypeptide or immunogenic peptide of the invention, or a nucleic acid expressing such a polypeptide or peptide, can be administered to an animal, using standard methods, and polyclonal antibodies isolated therefrom. Such polypeptides or peptides, if desired, can be conjugated to a carrier, such as KLH, serum albumin, tetanus toxoid and the like, using standard linking techniques, to increase their immunogenicity. Additionally, such peptides can be formulated together with an adjuvant known in the art, such as Freund's complete or incomplete adjuvant. The antibodies so generated can be used in the form of serum isolated from an immunized animal, or the antibody can be affinity purified from the serum using the invention peptides or polypeptides.

[0150] Additionally, the antibodies of the invention can be monoclonal antibodies produced by a hybridoma cell line, by chemical synthesis, or by recombinant methods. Modified antibodies, such as chimeric antibodies, single chain antibodies, humanized antibodies and CDR-grafted or bifunctional antibodies, can also be produced by methods well known to those skilled in the art.

[0151] Methods of preparing and using antibodies and antigen-binding fragments, including detectably labeled antibodies, are described, for example, in Harlow and Lane, Antibodies: A Laboratory Manual, Cold Spring Harbor Laboratory, New York (1989); in Day, E. D., Advanced Immunochemistry, Second Ed., Wiley-Liss, Inc., New York, N.Y. (1990); and in Borrebaeck (Ed.), Antibody Engineering, Second Ed., Oxford University Press, New York (1995). Example V describes a method for producing NR3B-specific antibodies.

[0152] The invention also provides a method for detecting an NR3B polypeptide in a sample. The method is practiced by contacting a sample with an antibody specific for an NR3B polypeptide and detecting specific binding of the antibody to the sample. As described above with respect to encoding nucleic acid molecules, altered expression, abundance or integrity of an NR3B polypeptide in a sample can thus be indicative of a pathological condition, including any of the acute or chronic neurological and neurodegenerative conditions described herein.

[0153] The methods of detecting an NR3B polypeptide in a sample can be either qualitative or quantitative, and can detect the presence, abundance, integrity or localization of the polypeptide, as desired for a particular application. Contemplated assays to detect a polypeptide in a sample include in situ histochemistry, immunoblotting, immunoprecipitation, FACS analysis, radioligand binding, and ELISA analysis. Such assays can be direct, using a detectably labeled ligand, or indirect, using a labeled secondary reagent, such as an anti-ligand antibody. Exemplary labels include fluorescent labels, enzymes, radioisotopes, and biotin. Detection can be by any convenient analytical means, including by spectrophotometric, radiographic or chemiluminescent means, depending on the assay.

[0154] The invention also provides a method of detecting an NR3B ligand. In one embodiment, the method is practiced by contacting an NR3B polypeptide with one or more compounds under conditions suitable for detecting binding to the polypeptide, and detecting a candidate compound that binds the polypeptide. In another embodiment, the invention is practiced by contacting an NR3B functional fragment with one or more compounds under conditions suitable for detecting binding to the fragment, and detecting a candidate compound that binds the fragment. For use in such an application, the NR3B functional fragment desirably contains at least one extracellular domain of the NR3B polypeptide, such as the region N-terminal to the S1 domain, the S1 domain, the S2 domain, or the region between the M3 and M4 domains (see FIG. 8).

[0155] The invention further provides a method of detecting an excitatory glycine receptor ligand. The method is practiced by contacting an excitatory glycine receptor with one or more compounds under conditions suitable for detecting binding to the receptor, and detecting a candidate compound that binds the receptor.

[0156] In one embodiment, the excitatory glycine receptor contains an NR3B polypeptide and an NR1 polypeptide. In another embodiment, the excitatory glycine receptor contains an NR3A polypeptide and an NR1 polypeptide. In a further embodiment, the excitatory glycine receptor contains an NR3B polypeptide, an NR3A polypeptide and an NR1 polypeptide. In yet another embodiment, the excitatory glycine receptor contains an NR3B and/or an NR3A polypeptide, an NR1 polypeptide and an NR2 polypeptide (i.e. an NR2A, 2B, 2C or 2D polypeptide).

[0157] As disclosed herein, co-expression of an NR3A and an NR1 polypeptide also results in the formation of excitatory glycine receptors. Rat NR3A polypeptides and encoding nucleic acid molecules are known in the art, and exemplary molecules have the nucleotide and predicted amino acid sequences set forth in Table 2: TABLE 2 SUBUNIT ACCESSION NUMBER Rat NR3A (NMDAR-L) U29873 Rat NR3A (x-1) L34938 Rat NR3A (splicing variant) AF061945 Rat NR3A (splicing variant) AF073379

[0158] Disclosed herein as SEQ ID NO:53 is the predicted human NR3A cDNA sequence and its encoding amino acid sequence (SEQ ID NO:54) (see FIGS. 11 and 12). These sequence were predicted from the human NR3A genomic sequences in the database (GenBank Accession Nos. NT_(—)025809, AL35616, XM042803.1 and AL359651.1) based on the rat NR3A cDNA and amino acid sequences. The skilled person can likewise determine additional NR3A sequences from other species.

[0159] NR2 polypeptides and encoding nucleic acid molecules are also known in the art, and exemplary molecules have the nucleotide and predicted amino acid sequences set forth in Table 3: TABLE 3 SUBUNIT ACCESSION NUMBER Human NR2A cDNA NM_000833, U09002, U90277, XM_030214, XM_030215, XM_007911, XM_016282 Human NR2A genomic AC007218, AC006531 Rat NR2A cDNA NM_012573, M91561, AF001423, D13211 Mouse NR2A (epsilon 1) NM_008170, D10217 Human NR2B cDNA XM_006636, XM_042404, XM_042405, XM_042406, NM_000834, U88963, U11287, U90278 Human NR2B genomic AC007535 Rat NR2B cDNA NM_012574, U11419, M91562 Mouse NR2B (epsilon 2) NM_008171, D10651 Human NR2C cDNA NM_000835, U77782, XM_046949, L76224, XM_012596 Rat NR2C NM_012575, D13212, M91563, U08259 Mouse NR2C (epsilon 3) NM_010350, D10694 Human NR2D cDNA XM_009108, NM_000836, U77783 Human NR2D genomic AC011527, AC008403 Rat NR2D U08260, NM_022797, L31612, L31611, D13213 Mouse NR2D (epsilon 4) NM_008172, D12822

[0160] The skilled person will appreciate that for use in the methods described herein, an NR3A or NR2 polypeptide can have modifications to a naturally occurring sequence, or be a functional fragment of the naturally occurring sequence, so long as the desired biological activity is retained. An exemplary modification of a naturally occurring NR3A or NR2 polypeptide that does not affect biological activity is the addition of an epitope tag to facilitate identification in procedures such as immunolocalization and immunoprecipitation.

[0161] NR3A biological activities include, for example, the ability to oligomerize with other NMDA subunits, including NR1; the ability to bind glycine; and the ability to form excitatory glycine receptors in association with NR1. NR2 biological activities include, for example, the ability to oligomerize with other NMDA subunits, including NR1; the ability to bind glutamate; and the ability to form excitatory glutamate receptors in association with NR1.

[0162] As described above with respect to NR3B and NR1 polypeptides and functional fragments, the skilled person can readily make NR3A and NR2 molecules with sequences that differ from the naturally occurring sequence and test such molecules to confirm that a desired NR3A or NR2 biological activity is retained.

[0163] As used herein, the term “ligand” refers to any biological or chemical compound that binds the recited polypeptide, fragment or receptor with high affinity. High affinity binding refers to binding with a Kd of less than about 10⁻³M, such as less than 10⁻⁵M, and often less than 10⁻⁷M. Glycine and NR3B antibodies are examples of ligands of NR3B.

[0164] An “NR3B ligand” or “excitatory glycine receptor ligand” can further be an agonist or antagonist of an excitatory glycine receptor, as described below, or can be a compound having little or no effect on excitatory glycine receptor biological activity. For example, a ligand without agonistic or antagonistic activity can be used to specifically target a diagnostic or therapeutic moiety to cells and tissues that express an excitatory glycine receptor. Thus, an identified ligand can be labeled with a detectable moiety, such as a radiolabel, fluorochrome, ferromagnetic substance, or luminescent substance, and used to detect normal or abnormal expression of an excitatory glycine receptor in an isolated sample or in in vivo diagnostic imaging procedures. Likewise, an identified ligand can be labeled with a therapeutic moiety, such as a cytotoxic or cytostatic agent or radioisotope, and administered in an effective amount to arrest proliferation or kill a cell or tissue that aberrantly expresses an excitatory glycine receptor for use in therapeutic applications described further below.

[0165] Binding assays, including high-throughput automated binding assays, are well known in the art and can be used in the invention methods. The assay format can employ a cell, cell membrane, artificial membrane system, or purified polypeptide, fragment or receptor, either in solution or attached to a solid phase. If desired, the binding assay can be performed in the presence of a known ligand of NR3B or of an excitatory glycine receptor, such as glycine.

[0166] Suitable assays that can be used for detecting ligand binding include, for example, scintillation proximity assays (SPA) (Alouani, Methods Mol. Biol. 138:135-41 (2000)), UV or chemical cross-linking (Fancy, Curr. Opin. Chem. Biol. 4:28-33 (2000)), competition binding assays (Yamamura et al., Methods in Neurotransmitter Receptor Analysis, Raven Press, New York, 1990), biomolecular interaction analysis (BIA) (Weinberger et al., Pharmacogenomics 1:395-416 (2000)), mass spectrometry (MS) (McLafferty et al., Science 284:1289-1290 (1999) and Degterev, et al., Nature Cell Biology 3:173-182 (2001)), nuclear magnetic resonance (NMR) (Shuker et al., Science 274:1531-1534 (1996), Hajduk et al., J. Med. Chem. 42:2315-2317 (1999), and Chen and Shapiro, Anal. Chem. 71:669A-675A (1999)), fluorescence polarization assays (FPA) (Degterev et al., supra, 2001); surface plasmon resonance (SPR)(Liparoto et al., J. Mol Recognit. 12:316-321 (1999)); and protein chip proteomic array analysis (e.g. ProteinChip™ System from Ciphergen Biosystems, which can be used in tandem with mass spectrometry analysis for sequence or structure determination).

[0167] An exemplary assay that has been used successfully to identify ligands of an NMDA receptor is phage display (see Li et al., Nature Biotech. 14:986-991 (1996), which describes contacting an N-terminal fragment of an NR1 polypeptide with a phage display library). A similar phage display approach can be applied to determine NR3B ligands and excitatory glycine receptor ligands.

[0168] Exemplary high-throughput receptor binding assays are described, for example, in Mellentin-Micelotti et al., Anal. Biochem. 272:P182-190 (1999); Zuck et al., Proc. Natl. Acad. Sci. USA 96:11122-11127 (1999); and Zhang et al., Anal. Biochem. 268;134-142 (1999). Other suitable methods are known in the art.

[0169] The invention also provides methods of detecting an excitatory glycine receptor agonist or antagonist The method is practiced by contacting an excitatory glycine receptor under conditions suitable for detecting excitatory glycine receptor activation, and detecting a candidate compound that alters excitatory glycine receptor activation. As described herein, excitatory glycine receptor activation can be evidenced by elicitation of a monovalent cation current, with little or no channel permeability to Ca²⁺ and little or no inhibition by Mg²⁺.

[0170] In one embodiment, the excitatory glycine receptor contains an NR3B polypeptide and an NR1 polypeptide. In another embodiment, the excitatory glycine receptor contains an NR3A polypeptide and an NR1 polypeptide. In a further embodiment, the excitatory glycine receptor contains an NR3B polypeptide, an NR3A polypeptide and an NR1 polypeptide. In yet another embodiment, the excitatory glycine receptor contains an NR3B polypeptide, an NR1 polypeptide and an NR2 polypeptide (i.e. an NR2A, 2B, 2C or 2D polypeptide). In another embodiment, the excitatory glycine receptor contains an NR3A polypeptide, an NR1 polypeptide and an NR2 polypeptide (i.e. an NR2A, 2B, 2C or 2D polypeptide).

[0171] The agonists and antagonists identified by the methods of the invention are useful in therapeutic applications, described further below, in which it is desirable to increase or decrease ion flow through the excitatory glycine receptor.

[0172] As used herein, the term “excitatory glycine receptor agonist” refers to a compound that increases or activates excitatory glycine receptor cation currents. An agonist can act by any mechanism, such as by binding the receptor at the normal glycine binding site, thereby mimicking glycine and promoting receptor activation. An agonist can also act, for example, by potentiating the activity of glycine, or by favorably altering the conformation of the receptor. The methods of the invention can be used to detect agonists that act through any agonistic mechanism.

[0173] As used herein, the term “excitatory glycine receptor antagonist” refers to a compound that decreases or inhibits excitatory glycine receptor cation currents. Typically, the effect of an antagonist is observed as a blocking of activation by an agonist. For example, 5, 7-di-Cl-Kynurenate blocks glycine activated currents through the NR3B/NR1 receptor.

[0174] Antagonists include, for example, partial antagonists, partial agonists, competitive antagonists, non-competitive antagonists and uncompetitive antagonists. A competitive antagonist interacts with or near the site specific for the agonist. A non-competitive antagonist inactivates the function of the receptor by interacting with a site other than the site that interacts with the agonist. Partial agonists have both agonistic and antagonistic activity. For example, as shown in FIG. 3, D-serine evokes small NR1/NR3B currents alone, but dose-dependently inhibits currents in the presence of glycine. The methods of the invention can be used to detect antagonists that act through any antagonistic mechanism.

[0175] Methods of detecting excitatory glycine receptor agonists and antagonists can advantageously be performed either in the presence or absence of a physiologically relevant excitatory glycine receptor agonist such as glycine. Compounds that demonstrate agonistic and antagonistic effects in the presence of glycine are particularly useful for use in therapeutic applications, in which physiological concentrations of circulatory glycine are present. In such methods, concentrations of glycine of about 1 to about 100 μM, such as about 5-10 μM, are suitable.

[0176] Electrophysiological methods for detecting monovalent cation currents through an excitatory glycine receptor are well known in the art. Exemplary methods for recording whole-cell and single-channel currents in Xenopus oocytes, brain slices, mammalian cells and cell-free membrane patches are described in Das et al., Nature 393:377-381 (1998); Sakmann and Neherand, in Single-Channel Recording, 2nd ed., Ch. 15, pp. 341-355, (1995), edited by Bert Sakmann and Erwin Neher, Plenum Press, New York; Penner, in Single-Channel Recording, 2nd ed., Ch. 1, pp. 3-28; Hamill et al., Pflugers Arch. 391:85-100 (1981); Ilers et al., in Single-Channel Recording, 2nd ed., Ch. 9, pp. 213-229, (1995), edited by Bert Sakmann and Erwin Neher, Plenum Press, New York; and in the Examples, below.

[0177] Ionic currents can also be detected using suitable detectably labeled ion indicators. Ion indicators and methods for their use are known in the art. For example, monovalent cation currents through the excitatory glycine receptor can be detected using Na⁺ or K⁺ ion indicators, which can be fluorescently labeled or radiolabeled (see, for example, Moore et al., Proc. Natl. Acad. Sci. USA 90:8058-8062 (1993); Paucek et al., J. Biol. Chem. 267:26062-26069 (1992); Xu et al., J. Biol. Chem. 270: 19606-19612 (1995)). Exemplary ion indicators include: SBFI sodium indicator, Sodium Green sodium indicator; CoroNa Red sodium indicator; PBFI potassium indicator; 6-Methoxy-N-(3-sulfopropyl)quinolinium (SPQ) chloride indicator; N-(Ethoxycarbonylmethyl)-6-methoxyquinolinium bromide (MQAE) chloride indicator; 6-Methoxy-N-ethylquinolinium iodide (MEQ) chloride indicator; Lucigenin chloride indicator, which are available from Molecular Probes, Inc.

[0178] Subsequent to excitatory glycine receptor activation and membrane depolarization, an influx of Ca²⁺ ions occurs if voltage-dependent Ca²⁺ channels are present in the cell being studied. If the cell of interest does not endogenously express voltage-dependent Ca²⁺ channels, the cell can be recombinantly engineered to express such channels, using voltage-dependent Ca²⁺ channel subunit gene sequences and molecular biology methods known in the art. Accordingly, ionic currents through the excitatory glycine receptor can also be detected, indirectly, using detectably labeled Ca²⁺ ion indicators, which can be fluorescently labeled or radiolabeled. Exemplary Ca²⁺ ion indicators include FLUO-3 AM, FLUO-4 AM, FURA-2, INDO-1, FURA RED, CALCIUM GREEN, CALCIUM ORANGE, CALCIUM CRIMSON, BTC, and OREGON GREEN BAPTA (see, for example, Grynkiewitz et al., J. Biol. Chem. 260:3440-3450 (1985); Sullivan et al., in Calcium Signal Protocol, Methods in Molecular Biology 114: 125-133, Edited by David G. Lambert, Human Press, Totowa, N.J. (1999); Miyawaki et al., Proc. Natl. Acad. Sci. USA 96:2135-2140 (1999); and Coward et al., Analyt. Biochem. 270:242-248 (1999)).

[0179] Assay methods for identifying compounds that bind to or modulate excitatory glycine receptor activity (e.g. ligands, agonists and antagonists) generally involve comparison to a control. One type of a “control” is a NR3B polypeptide or excitatory glycine receptor that is treated substantially the same as the polypeptide or receptor exposed to the candidate compound, except the control is not exposed to the candidate compound. For example, the same recombinant cell can be tested in the presence and absence of candidate compound, by merely changing the solution contacting the cell. Another type of “control” is a cell that is essentially identical to the NR3B polypeptide- or excitatory glycine receptor-expressing recombinant cell, except the control cell does not express the polypeptide or receptor. In this situation, the response of the test cell to a candidate compound is compared to the response (or lack of response) of the control cell to the same compound under substantially the same reaction conditions.

[0180] The term “candidate compound” refers to any molecule that potentially acts as a ligand, agonist or antagonist or ligand in the screening methods disclosed herein. A candidate compound can be a naturally occurring macromolecule, such as a polypeptide, amino acid, nucleic acid, carbohydrate, lipid, or any combination thereof. A candidate compound also can be a partially or completely synthetic derivative, analog or mimetic of such a macromolecule, or a small organic molecule prepared by combinatorial chemistry methods. If desired in a particular assay format, a candidate compound can be detectably labeled or attached to a solid support.

[0181] Methods for preparing large libraries of compounds, including simple or complex organic molecules, metal-containing compounds, carbohydrates, peptides, proteins, peptidomimetics, glycoproteins, lipoproteins, nucleic acids, antibodies, and the like, are well known in the art and are described, for example, in Huse, U.S. Pat. No. 5,264,563; Francis et al., Curr. Opin. Chem. Biol. 2:422-428 (1998); Tietze et al., Curr. Biol., 2:363-371 (1998); Sofia, Mol. Divers. 3:75-94 (1998); Eichler et al., Med. Res. Rev. 15:481-496 (1995); and the like. Libraries containing large numbers of natural and synthetic compounds also can be obtained from commercial sources.

[0182] The number of different candidate compounds to test in the methods of the invention will depend on the application of the method. For example, one or a small number of candidate compounds can be advantageous in manual screening procedures, or when it is desired to compare efficacy among several predicted ligands, agonists or antagonists. However, it is generally understood that the larger the number of candidate compounds, the greater the likelihood of identifying a compound having the desired activity in a screening assay. Additionally, large numbers of compounds can be processed in high-throughput automated screening assays. Therefore, “one or more candidate compounds” can be, for example, 2 or more, such as 5, 10, 15, 20, 50 or 100 or more different compounds, such as greater than about 10³, 10⁵ or 10⁷ different compounds, which can be assayed simultaneously or sequentially.

[0183] The function of the NR3A polypeptide has remained elusive, despite years of investigation (see Ciaberra et al., J. Neuroscience 15:6498-6508 (1995); Sucher et al., J. Neuroscience 15:6509-6520 (1995); Das et al. supra (1998); Perez-Otano et al., J. Neuroscience 21:1228-1237 (2001)). As disclosed herein, when expressed in combination with NR1 and optionally also NR2 subunits, the NR3A polypeptide, like the NR3B polypeptide, forms an excitatory glycine receptor (see Example IV). The invention thus provides isolated excitatory glycine receptors containing either an NR3A polypeptide or an NR3B polypeptide, or both. Such receptors are suitable use in the methods described herein of detecting excitatory glycine receptors agonists, antagonists and ligands. As described previously, depending on the particular assay, the isolated receptors can optionally be present at the surface of an intact cell, present in a cell membrane with or without other cellular components, or reconstituted into a natural or artificial lipid bilayer.

[0184] In one embodiment, the excitatory glycine receptor contains an NR3B polypeptide and an NR1 polypeptide. In another embodiment, the excitatory glycine receptor contains an NR3A polypeptide and an NR1 polypeptide. In a further embodiment, the excitatory glycine receptor contains an NR3B polypeptide, an NR3A polypeptide and an NR1 polypeptide. In yet another embodiment, the excitatory glycine receptor contains an NR3B polypeptide, an NR1 polypeptide and an NR2 polypeptide (i.e. an NR2A, 2B, 2C or 2D polypeptide). In another embodiment, the excitatory glycine receptor contains an NR3A polypeptide, an NR1 polypeptide and an NR2 polypeptide (i.e. an NR2A, 2B, 2C or 2D polypeptide).

[0185] Suitable NR3B, NR3A, NR2A-D and NR1 polypeptides that can be used as subunits of excitatory glycine receptors, including both naturally occurring polypeptides, and modifications and functional fragments of such polypeptides, have been described previously.

[0186] The invention also provides therapeutic methods for the prevention and amelioration of conditions in which inappropriate NMDA receptor activation, or inappropriate responses to glycine or glutamate, are implicated. Such conditions include, for example, acute neurologic condition, such as cerebral ischemia; stroke; hypoxia; anoxia; poisoning by carbon monoxide, manganese, cyanide or domoic acid; hypoglycemia; mechanical trauma to the nervous system such as trauma to the head or spinal cord; or epileptic seizure. Other conditions include, for example, chronic neurodegenerative disease, such as Huntington's disease; a disorder of photoreceptor degeneration such as retinitis pigmentosa; acquired immunodeficiency syndrome (AIDS) dementia complex (HIV-associated dementia); a neuropathic pain syndrome such as causalgia or a painful peripheral neuropathy; olivopontocerebellar atrophy; Parkinsonism; amyotrophic lateral sclerosis; a mitochondrial abnormality or other biochemical disorder such as MELAS syndrome, MERRF, Leber's disease, Wernicke's encephalopathy, Rett syndrome, homocysteinuria, hyperhomocysteinemia, hyperprolinemia, nonketotic hyperglycinemia, hydroxybutyric aminoaciduria, sulfite oxidase deficiency, combined systems disease, lead encephalopathy, Alzheimer's disease, hepatic encephalopathy, Tourette's syndrome, drug addiction/tolerance/dependency, glaucoma, depression, anxiety, multiple sclerosis and other demyelinating disorders. Other conditions are known in the art and reviewed, for example, in Lipton et al., New Engl. J. Med. 330:613-622 (1994) and Cull-Candy et al., Curr. Opin. Neurobiol. 11:327-335 (2001).

[0187] Thus, the invention provides methods for increasing or decreasing signaling through an excitatory glycine receptor by administering an excitatory glycine receptor agonist, antagonist or ligand, or an NR3B ligand, to an individual. Methods of identifying such agonist, antagonist and ligands have been described previously.

[0188] The invention also provides gene therapy methods for modulating a cellular response to glycine or glutamate. By overexpressing a full-length NR3B polypeptide, the NR1 subunit can form NR1/NR3B receptors, which are insensitve to glutamate, rather than NR1/NR2 receptors. Therefore, detrimental glutamate responses can be reduced. In contrast, by expressing a dominant negative NR3B polypeptide, or a construct that prevents translation of endogenous NR3B, fewer functional NR1/NR3B receptors will be formed. Therefore, detrimental glycine responses can be reduced. Therapeutic applications in which it is desirable to modulate cellular responses to glutamate and glycine are described above.

[0189] In one embodiment, the invention provides a method of modulating a cellular response to glycine or glutamate by introducing a nucleic acid molecule encoding an NR3B polypeptide or functional fragment into a cell, and expressing the NR3B functional fragment encoded by the nucleic acid molecule in the cell. In another embodiment, the invention provides a method of modulating a cellular response to glycine or glutamate by introducing an antisense nucleic acid molecule, a ribozyme molecule or a small interfering RNA (siRNA) molecule into the cell, wherein the molecule hybridizes to an NR3B nucleic acid molecule and prevents translation of the encoded NR3B polypeptide.

[0190] Suitable gene therapy vectors have been described previously. For gene therapy applications, the nucleic acid molecule can be administered to a subject by various routes. For example, local administration at the site of a pathology can be advantageous because there is no dilution effect and, therefore, the likelihood that a majority of the targeted cells will be contacted with the nucleic acid molecule is increased. This is particularly true in the eye, where either intravitreal or intraretinal administration is possible. In addition, administration can be systemic, such as via intravenous or subcutaneous injection into the subject. For example, following injection, viral vectors will circulate until they recognize host cells with the appropriate target specificity for infection.

[0191] Receptor-mediated DNA delivery approaches also can be used to deliver a nucleic acid molecule into cells in a tissue-specific manner using a tissue-specific ligand or an antibody that is non-covalently complexed with the nucleic acid molecule via a bridging molecule. Direct injection of a naked nucleic acid molecule or a nucleic acid molecule encapsulated, for example, in cationic liposomes also can be used for stable gene transfer into non-dividing or dividing cells. In addition, a nucleic acid molecule can be transferred into a variety of tissues using the particle bombardment method.

[0192] Antisense nucleotide sequences that are complementary to a nucleic acid molecule encoding an NR3B polypeptide can be used to prevent or reduce NR3B expression. Therefore, the method can be practiced with an antisense nucleic acid molecule complementary to at least a portion of the nucleotide sequence of SEQ ID NOS:1, 3, 5 or 7. For example, the antisense nucleic acid molecule can be complementary to a region within the N-terminus of SEQ ID NOS:1, 3, 5 or 7, such as within nucleotides 1-1000, 1-500, 1-100 or 1-18, and can optionally include sequences 5′ to the start codon. Methods of preparing antisense nucleic acids molecules and using them therapeutically are known in the art and described, for example, in Galderisi et al., J. Cell Physiol. 181:251-257 (1999).

[0193] Likewise, ribozymes that bind to and cleave SEQ ID NOS:1, 3, 5 or 7 can also be effective in preventing or reducing NR3B expression. Methods of preparing ribozymes and DNA encoding ribozymes, including hairpin and hammerhead ribozymes, and using them therapeutically are known in the art and described, for example, in Lewin et al., Trends Mol. Med. 7:221-228 (2001).

[0194] Additionally, small interfering RNAs (siRNAs), which are short duplex RNAs with overhanging 3′ ends, directed against SEQ ID NOS:1, 3, 5 or 7, can also be effective in preventing or reducing NR3B expression. Methods of preparing and using siRNAs are known in the art and described, for example, in Elbashir et al., Nature 411:494-498 (2001).

[0195] The therapeutic compounds of the invention, including agonists, antagonists, ligands and nucleic acid molecules, can be formulated and administered in a manner and in an amount appropriate for the condition to be treated; the weight, gender, age and health of the individual; the biochemical nature, bioactivity, bioavailability and side effects of the particular compound; and in a manner compatible with concurrent treatment regimens. An appropriate amount and formulation for a particular therapeutic application in humans can be extrapolated based on the activity of the compound in the in vitro binding and signaling assays described herein, or from recognized animal models of the particular disorder.

[0196] The total amount of therapeutic compound can be administered as a single dose or by infusion over a relatively short period of time, or can be administered in multiple doses administered over a more prolonged period of time. Additionally, the compound can be administered in a slow-release matrice, which can be implanted for systemic delivery at or near the site of the target tissue. Contemplated matrices useful for controlled release of therapeutic compounds are well known in the art, and include materials such as DepoFoam™, biopolymers, micropumps, and the like.

[0197] The therapeutic compounds can be administered to an individual by routes known in the art including, for example, intravenously, intramuscularly, subcutaneously, intraorbitally, intracapsularly, intraperitoneally, intracisternally, intra-articularly, intracerebrally, orally, intravaginally, rectally, topically, intranasally, transdermally or intravitreally.

[0198] Preferably, the therapeutic compounds are administered to a subject as a pharmaceutical composition comprising the compound and a pharmaceutically acceptable carrier. The choice of pharmaceutically acceptable carrier depends on the route of administration of the compound and on its particular physical and chemical characteristics. Pharmaceutically acceptable carriers are well known in the art and include sterile aqueous solvents such as physiologically buffered saline, and other solvents or vehicles such as glycols, glycerol, oils such as olive oil and injectable organic esters. A pharmaceutically acceptable carrier can further contain physiologically acceptable compounds that stabilize the compound, increase its solubility, or increase its absorption. Such physiologically acceptable compounds include carbohydrates such as glucose, sucrose or dextrans; antioxidants, such as ascorbic acid or glutathione; chelating agents; and low molecular weight proteins.

[0199] For applications that require the compounds and compositions to cross the blood-brain barrier, or to cross the cell membrane, formulations that increase the lipophilicity of the compound are particularly desirable. For example, the compounds of the invention can be incorporated into liposomes (Gregoriadis, Liposome Technology, Vols. I to III, 2nd ed. (CRC Press, Boca Raton Fla. (1993)). Liposomes, which consist of phospholipids or other lipids, are nontoxic, physiologically acceptable and metabolizable carriers that are relatively simple to make and administer. Additionally, the therapeutic compound can be conjugated to a peptide that facilitates cell entry, such as penetratin (also known as Antennapedia peptide), other homeodomain sequences, or the HIV protein Tat.

[0200] It is contemplated that administration of the therapeutic compounds and compositions of the invention will result in some beneficial effect to the individual, such as improved overall neurological function or a specific neurological function; an improvement in the quality of life; a reduction in the severity of the symptoms of the disease; a reduction in the number of diseased cells; prolonged survival, and the like. Indicators of beneficial effect are well known in the art, and an appropriate indicator for a particular application can be determined by the skilled person.

[0201] The following examples are intended to illustrate but not limit the present invention.

EXAMPLE I

[0202] This example shows cloning and sequence analysis of NR3B.

[0203] Degenerate primers were designed based on the sequences of NR3A and other NMDAR family members. Using degenerate PCR in concert with homology screening of a rat brain cDNA library, two novel cDNA fragments (called 5-2 clone 1 and 5-2 clone 2) were obtained which exhibited significant sequence identity with NR3A, but clearly corresponded to a distinct and previously unidentified gene. The novel gene was therefore designated NR3B.

[0204] Based on the sequences of 5-2 clone 1 and 5-2 clone 2, two pairs of more specific, nested primers (f4, TGCTGCTATGGCTACTGCATC (SEQ ID NO:17); r4, ATGACAGCAGCCAGGTTGGCCGT (SEQ ID NO:18); f5, CACACATGGCTGTGACCAGC (SEQ ID NO:19); and r5, AGAATGGCATAGCACAGGTTG (SEQ ID NO:20)) were designed and used to PCR screen a Rapid-Screen rat brain cDNA library panel (OriGene Technologies, Inc., Rockville, Md.). Four independent partial NR3B cDNA clones were thus obtained. The coding regions of three of the clones were identical (one of these was designated clone B4), but the fourth clone (clone A2) had a 45-nucleotide deletion in the 5′ coding region, which may, therefore, represent a splice variant of NR3B. A comparison of the sequences of clones B4, A2, 5-2 clone 1, and 5-2 clone 2 revealed that all overlap, e.g., 5-2 clone 2 spans 49% of the 5′ end, and clones B4 and A2 span 92% of the 3′ end of the full-length NR3B cDNA. Full-length NR3B cDNAs were assembled by ligating the EcoRI-XhoI fragment of 5-2 clone 2 (345 bp) to the XhoI-XbaI fragment of either clone B4 (2847 bp) or clone A2 (2802 bp).

[0205] The NR3B (B4 form) cDNA has an open reading frame encoding a peptide of 1003 amino acid (aa) residues. Multiple alignments of the predicted amino acid sequence of NR3B with other glutamate receptor subunits indicated that NR3B is closely related to NR3A (47% identity), but shares less identity with the NR1 and NR2 subfamilies (17 to 21%) (FIG. 1). To a lesser degree, NR3B also shares sequence identity (13 to 16%) with other GluR subunits representing non-NMDAR subunits.

[0206] The major structural features of NR3A are highly conserved in NR3B, including the S1 and S2 agonist binding domains, and the membrane spanning regions (FIG. 1). A detailed comparison of the critical residues involved in ligand binding to the NR1 and NR2 subunits revealed that NR3B could potentially share ligand specificity with either or both of these classes of NMDA receptor subunits. Five of the seven amino acid residues required for glycine binding to NR1 and six of the ten amino acid residues required for glutamate binding to NR2A are conserved in NR3B. Furthermore, the M2 region of NR3B is similar to that of NR3A, but strikingly different from those of the NR1 and NR2 subunits. The M2 region forms the re-entrant P-loop which lines the ion channel pore of fully assembled NMDA receptors. These molecular features suggest that the NR3B subunit may confer novel ligand binding and channel gating properties to the NMDA receptor complexes into which it is assembled.

[0207] The first 500 aa residues in the extracellular N-terminus and intracellular C-terminus are the most divergent regions between NR3B and other NMDA receptor subunits, including the NR3A subunit. These regions are important for intra-subunit assembly of glutamate receptors and for interaction of the receptor complex with associated proteins. Thus, these differences may uniquely specify the selection of partner subunits, subcellular localization and CNS distribution of the NR3B subunit.

[0208] Based on the rat NR3B sequence, the corresponding mouse and human NR3B sequences were identified from an analysis of homologous genomic sequences in the database (Accession numbers AC087114 and AC004528, respectively).

EXAMPLE II

[0209] This example shows analysis of the expression and localization of the NR3B subunit.

[0210] Initially, in situ hybridization was used to detect NR3B mRNA in adult rat brain. Three probes of approximately 400 bp were generated from different N-terminal and C-terminal regions of NR3B. These regions shared little sequence identity with other NMDAR subunits, including NR3A. All three probes detected NR3B signals in the initial experiment. The probe in the N-terminal region giving the strongest signal was chosen for subsequent experiments. ³³P-labeled antisense probes detected “hot spots” in the facial and trigeminal nuclei of the brainstem, and in the ventral horn of the spinal cord (FIGS. 2a, b). Subsequent use of non-isotopic (digoxigenin) labeled probes yielded signals of much higher resolution at the cellular level. NR3B was not detectable in most brain areas, including the cerebrocortex, thalamus, basal ganglia, and hippocampus. In contrast, the spinal cord and the brainstem displayed very strong NR3B signals. In the spinal cord, the NR3B signal was found in motor neurons of the anterior horn at all levels in both layers VIII and IX, which innervate proximal and distal muscle groups, respectively (FIGS. 2c, d). However, an NR3B signal was detected only in specific motor nuclei of the brainstem, specifically the facial and trigeminal nuclei. In addition, some large cells in the vestibular nucleus and reticular formation were also positive for NR3B mRNA.

[0211] Further analysis using rats at different ages (P2, P7, P14 and adult) confirmed that NR3B mRNA expression begins at an early postnatal stage (FIG. 2d). A weak NR3B signal was detectable at P2 but increased substantially by P7. The level of NR3B mRNA expression increased postnatally, reaching a peak at P14, and remained elevated into adulthood, consistent with a specific role of NR3B in developing and maintaining motor neuron functions. Based upon these data, it was concluded that NR3B mRNA is localized mainly in motor neurons of the spinal cord and in some motor nuclei of the brainstem. Expression of NR3B protein in these regions has been confirmed by immunocytochemistry.

EXAMPLE III

[0212] This example shows characterization of biological activities of NR3B-containing excitatory glycine receptors.

[0213] NR3B cRNA for oocyte expression was produced as follows. The full-length NR3B cDNA was constructed in the pCMV6-XL4 vector (OriGene Technologies, Inc., Rockville, Md.), which contains a T7 promoter upstream of the cDNA insert. Initial attempts to generate an NR3B cRNA using T7 RNA polymerase resulted in the formation of both full-length and truncated species. The truncated cRNA could potentially encode a dominant negative form of NR3B. Therefore, a new vector was constructed to facilitate production of exclusively full-length NR3B cRNA. Construction of this vector proceeded as follows. NR1 cDNA was excised from the pGEM-HE/NR1 vector (a gift from S. F. Heinemann) by digestion with EcoRI and the two ends of the truncated pGEM-HE vector were re-ligated. The truncated pGEM-HE vector retained the 5′ and 3′ UTRs of the X. laevis β-globin gene for high level protein expression in frog oocytes. The SfoI-KpnI fragment of truncated pGEM-HE, which contains the T7 promoter, was replaced with the PvuII-KpnI fragment of pBluescript II KS, which contains the T3 promoter, to generate pJC32. The abbreviated multiple cloning site of pJC32 was then replaced with the multiple cloning site from pcDNA3.1(+) (Invitrogen, Carlsbad, Calif.) by ligating the 102 bp PmeI fragment of pcDNA3.1(+) into the 3 kb SmaI-EcoRV fragment of pJC32 to generate pJC34. Oligos encoding restriction sites for AscI, PacI, and PmeI (ctagcGGCGCGCCTTAATTAAGTTTAAACg (SEQ ID NO:52) and ctagcGTTTAAACTTAATTAAGGCGCGCCg (SEQ ID NO:53)) were annealed and ligated into the NheI site of pJC34. The resulting vector, pJC39, contains three rare restriction sites downstream of the multiple cloning site to facilitate linearization prior to in vitro transcription using T3 RNA polymerase.

[0214] The full-length NR3B (B4 form) cDNA was then excised from pCMV6-XL4 by digestion with EcoRI and XbaI, and the 3.2 kb fragment was ligated into EcoRI/XbaI-digested pJC39. The resulting vector, pJC42, was linearized with PmeI, and in vitro transcription was performed using the mMessage mMachine T3 kit (Ambion, Inc., Austin, Tex.) according to the manufacturer's instructions. The cRNA product consisted of a single species with a molecular weight of approximately 1.5 kb.

[0215] Microinjection of oocytes and electrophysiological recordings were performed as follows. Physiological and pharmacological properties of NMDA receptor subunits were characterized in the X. laevis oocyte expression system. Full-length cDNAs encoding NMDA receptor subunits were linearized and used as templates for synthesis of cRNA by in vitro transcription using T3 or T7 RNA polymerase (Ambion, Inc., Austin, Tex.). The DNA template was removed by DNase I digestion, and the RNA was purified by phenol/chloroform extraction and ethanol precipitation. The RNA was resuspended in water at 1 μg/μl and stored in −80° C. freezer until use. X. laevis oocytes were harvested and defolliculated by digestion with collagenase (2 mg/ml) for 2 hrs. Twenty to 40 ng of cRNA were injected into each oocyte using glass pipettes with a tip size of 20-40 μm in diameter. Electrophysiological recordings were performed from whole oocytes 2-7 days after injection using a dual-electrode voltage-clamp amplifier (Model OC-725A, Warner Instrument, Hamden, Conn.). Micropipettes were filled with 3M KCl. The bath solution contained 115 mM NaCl, 2.5 mM KCl, 1.5 mM BaCl₂, and 10 mM Hepes at pH 7.4. Drugs were applied to oocytes via a rapid perfusion system. Data were collected at 2.2 to 11.1 Hz at room temperature using the MacLab/4e A/D converter (AD Instruments, Mountain View, Calif.). Data analysis was performed using the programs Chart (AD Instruments, Mountain View, Calif.) and Microsoft Excel.

[0216] Additionally, using the patch-clamp technique, outside-out patches were pulled from oocytes previously injected with NMDAR cRNAs. This method allowed the recording of NMDA-evoked or glycine-evoked single-channel currents, as described previously (Das et al., supra (1998)).

[0217] Several novel properties of the NR3B subunit were identified by functional expression of the B4 form in Xenopus oocytes. First, functional receptors (defined by the presence of glycine-evoked current) were assembled in oocytes after co-injection of cRNAs for the NR3B and NR1 subunits. Functional receptors were also observed after co-injection of NR3B cRNA with both NR1 and NR2A subunit cRNAs. However, oocytes expressing NR1/NR2A/NR3B receptors exhibited ligand-evoked currents that were 5 to 10 times larger than those expressing NR1/NR3B receptors. In contrast, no functional receptors were detected when NR3B cRNA was co-injected with either NR2A or NR3A alone in the absence of NR1. These results suggest that assembly of the NR3R subunit into a functional receptor complex requires association with the NR1 subunit.

[0218] Strikingly, unlike the previously published NMDARs, it was found that the NR1/NR3B receptor was activated by glycine alone, in the absence of glutamate or NMDA (FIG. 3a). D-serine, D-alanine, D-cycloserine and ACPC, all co-agonists of traditional NMDARs via activation of the glycine site on the NR1 subunit, each inhibited glycine-induced currents of NR1/NR3B receptors (FIGS. 3b 1, b 2, and b 3). Importantly, NMDA, the traditional agonist for which this subclass of receptor was originally named, displayed little or no effect by itself or on the glycine-induced current of NR1/NR3B receptors (FIG. 3c). Aspartate and cysteine, previously shown to activate the glutamate binding site of NMDARs (containing NR2 subunits), slightly potentiated glycine-evoked currents of NR1/NR3B receptors, as did kainate which is known to activate non-NMDA (GluR5-7 and KA1,KA2) receptors (FIGS. 3d 1, 3 d 2). Of note, glycine alone activated NR1/NR3B receptors with high efficacy despite the fact that by itself glycine fails to excite the previously studied NMDARs or any other type of glutamate receptors. For NR1/NR3B receptors, the EC₅₀ for glycine was estimated to be approximately 5 μM. A more precise calculation of EC₅₀ was precluded by the desensitization that occurs at high glycine concentrations (FIG. 3a).

[0219] The altered ligand selectivity conferred by the NR3B subunit was demonstrated further by the totally unexpected effect of some NMDAR antagonists. For example, 2-amino-5-phosphonovalerate (APV), which normally acts as a competitive antagonist at the glutamate binding site of the known NMDARs, e.g., on NR2A in NR1/NR2A receptors, had little or no effect at 10 μM on glycine (10 μM)-evoked currents in NR1/NR3B receptors (FIG. 3f 1); at higher concentrations (40 μM), APV actually potentiated glycine-evoked current somewhat (right-hand panel, FIG. 4a). In contrast, 5,7-dichlorokynurenate, a competitive antagonist of the NR1 subunit glycine binding site, completely abolished glycine-evoked currents of NR1/NR3B receptors. As expected, strychnine, an antagonist of the inhibitory, chloride permeable glycine receptor, had no effect (FIG. 3f 2). Properties linked to the channel region were also altered by the presence of the NR3B subunit. Compared to NR1/NR2A receptors, NR1/NR3B receptors were drastically less sensitive to channel blockers such as Mg²⁺, MK-801, and memantine (FIGS. 3e 1, e 2, and e 3).

[0220] Like the more traditional NMDARs, it was found that NR1/NR3B receptor-operated channels were selectively permeable to cations. Replacement of all cations with N-methyl-D-gluconate (NMDG) in the external buffer completely abolished the glycine-induced inward current (FIG. 4a). Furthermore, replacement of all cations with Ca²⁺ also eliminated the inward current, and, unlike NR1/NR2 receptors, activation of NR1/NR3B receptors did not evoke a Ca²⁺-triggered Cl⁻ current when recorded in Ca²⁺-containing medium from Xenopus oocytes (data not shown). These results suggest that NR1/NR3B receptors display far less permeability to Ca²⁺ than NMDAR containing NR1 and NR2A-D subunits. Single-channel recording of NR1/NR3B receptors confirmed many of the aforementioned unique properties of these receptors, including activation by glycine alone and reduced sensitivity to Mg²⁺ (FIG. 5a). Recorded from outside-out patches of oocyte membrane, NR1/NR3B receptor-operated channels manifest a unitary conductance of ˜38 pS and a subconductance of 12 pS in the presence of 2 mM external Ba²⁺ (FIGS. 5a, c). When all anions in the patch electrode were replaced by impermeant gluconate, the conductance states remained unaffected at both positive and negative holding potentials (FIG. 5c), confirming that the channels were permeable to cations.

EXAMPLE IV

[0221] This example shows characterization of biological activities of NR3A-containing excitatory glycine receptors.

[0222] Because of the high degree of sequence similarity between NR3B and NR3A subunits, the ability of the NR3A subunit to form functional channels when co-expressed with NR1 was re-examined. Reminiscent of NR1/NR3B receptors, it was found that under specific conditions during two-electrode voltage recordings from oocytes, NR1/NR3A receptors could be activated by glycine alone (FIG. 6a). However, rapid desensitization of the current from NR1/NR3A receptors at micromolar concentrations of ligand rendered the observation of glycine-evoked currents extremely difficult, explaining why they had not been reported previously (Ciabarra et al., J. Neurosci. 15:6498-6508 (1995); Sucher et al., J. Neuroscience 15:6509-6520 (1995); Das et al., Nature 393:377-381 (1998).

[0223] Similar to NR1/NR3B receptors, glycine-evoked currents of NR1/NR3A receptors were effectively blocked by D-serine (FIG. 6b). APV blocked the current only slightly (FIG. 6c). As in the case of NR1/NR3B receptors, glycine-evoked currents from NR1/NR3A receptors were blocked by 5,7-dichlorokynurenate (FIG. 6d), but not by strychnine (FIG. 6e). Interestingly, channel blockers, such as Mg²⁺ and MK-801, not only failed to block glycine-evoked currents of NR1/NR3A receptors, but actually potentiated these currents when low concentrations of glycine were used (FIGS. 7a, b). These findings suggest that NR3A-containing receptors, similar to their NR3B counterparts, manifest novel pharmacological and physiological properties. Additionally, NR1/NR3A receptors desensitize rapidly, distinguishing them from NR1/NR3B receptors.

EXAMPLE V

[0224] This example shows the production of NR3B-specific antibodies using NR3B peptides.

[0225] In order to produce NR3B-specific antibodies, the XhoI-AvrII and MluI-XbaI fragments of rat NR3B cDNA (B4 form), which encode amino acids 89-238 and 927-1002, respectively, were subcloned into pET28 to facilitate expression of 6xHis-tagged NR3B fragments in bacteria. These regions of NR3B exhibit relatively poor homology with NR3A and other NMDA receptor subunits. The 6xHis-tagged proteins were expressed in BL21(DE3) cells and purified by metal affinity chromatography (Talon Purification Kit, Clontech, Palo Alto, Calif.). The purified proteins are used to immunize chickens (Aves Labs, Tigard, Oreg.) and rabbits (Covance, Princton, N.J.) for the production of anti-NR3B polyclonal antibodies.

[0226] All journal article, reference, sequence and patent citations provided above, in parentheses or otherwise, whether previously stated or not, are incorporated herein by reference in their entirety.

[0227] Although the invention has been described with reference to the examples provided above, it should be understood that various modifications can be made without departing from the spirit of the invention.

0 SEQUENCE LISTING <160> NUMBER OF SEQ ID NOS: 53 <210> SEQ ID NO 1 <211> LENGTH: 3133 <212> TYPE: DNA <213> ORGANISM: Rattus sp. <220> FEATURE: <221> NAME/KEY: CDS <222> LOCATION: (76)...(3036) <400> SEQUENCE: 1 gcacgagggg acaagagcgg gtctggctgg ggtgtctcct tcccttcgca cagcacagtg 60 gtaacttctt tcggg atg gag agt gtg cgg acg ctg tgg ctc agc gtg gcc 111 Met Glu Ser Val Arg Thr Leu Trp Leu Ser Val Ala 1 5 10 ctg gcg ctg gcg gtg ggg tcc cga gtg gtg cgc ggt cac cct cag ccc 159 Leu Ala Leu Ala Val Gly Ser Arg Val Val Arg Gly His Pro Gln Pro 15 20 25 tgc cgg gtt ccc acg cgc gct ggg gcc tcc gtg cgc ctg gcg gcg ctc 207 Cys Arg Val Pro Thr Arg Ala Gly Ala Ser Val Arg Leu Ala Ala Leu 30 35 40 ctg ccc cgg gcg ccc gcc gcc cgc gcc cgc gtc cta gct gcc ctg gcc 255 Leu Pro Arg Ala Pro Ala Ala Arg Ala Arg Val Leu Ala Ala Leu Ala 45 50 55 60 acc cct gcg ccg cgg ctg ccg cac aac ctg agt ctg gaa ctg gtg gcc 303 Thr Pro Ala Pro Arg Leu Pro His Asn Leu Ser Leu Glu Leu Val Ala 65 70 75 gtc gcg tcc ccg acc cgg gac ccc gcg tcg cta gct cga ggt ctg tgc 351 Val Ala Ser Pro Thr Arg Asp Pro Ala Ser Leu Ala Arg Gly Leu Cys 80 85 90 cag gtt ctg gca ccg cct ggc gtg gtg gcc tct ata gcc ttt ccc gag 399 Gln Val Leu Ala Pro Pro Gly Val Val Ala Ser Ile Ala Phe Pro Glu 95 100 105 gcg cgg ccc gag ctg cgg cta ctg cag ttc ctg gca gcc gcc aca gag 447 Ala Arg Pro Glu Leu Arg Leu Leu Gln Phe Leu Ala Ala Ala Thr Glu 110 115 120 acc cca gtg act ccg ttc cat ctg cag ctg gac tgg gct agt ccc ctg 495 Thr Pro Val Thr Pro Phe His Leu Gln Leu Asp Trp Ala Ser Pro Leu 125 130 135 140 gag acc ata ctg gat gtg ctg gtg tcc ctg gta cgg gca cat gcc tgg 543 Glu Thr Ile Leu Asp Val Leu Val Ser Leu Val Arg Ala His Ala Trp 145 150 155 gag gac att gct cta gta ctc tgc cgt gtc cgg gac cct ggc agc ctg 591 Glu Asp Ile Ala Leu Val Leu Cys Arg Val Arg Asp Pro Gly Ser Leu 160 165 170 gtg aca ctc tgg act aac cat gct agc cag gct cca aag ttt gtg ctg 639 Val Thr Leu Trp Thr Asn His Ala Ser Gln Ala Pro Lys Phe Val Leu 175 180 185 gac ctg agc cgg ctg gac agc agg aat gac agc ctt cgg gct gga ctg 687 Asp Leu Ser Arg Leu Asp Ser Arg Asn Asp Ser Leu Arg Ala Gly Leu 190 195 200 gcc ctg ttg ggg gcg ctg gaa gga ggg gga acc cca gtg cct gca gca 735 Ala Leu Leu Gly Ala Leu Glu Gly Gly Gly Thr Pro Val Pro Ala Ala 205 210 215 220 gtc ctc cta ggc tgc agc act gcc cgt gca cat gag gtc cta gag gct 783 Val Leu Leu Gly Cys Ser Thr Ala Arg Ala His Glu Val Leu Glu Ala 225 230 235 gca cca ccg ggt ccc cag tgg ttg ctg ggc aca cca ttg ccc gct gag 831 Ala Pro Pro Gly Pro Gln Trp Leu Leu Gly Thr Pro Leu Pro Ala Glu 240 245 250 gca ctg ccc acg act ggt ctg cca cct ggc gtg ctg gcg ctg ggg gaa 879 Ala Leu Pro Thr Thr Gly Leu Pro Pro Gly Val Leu Ala Leu Gly Glu 255 260 265 acc gaa caa cac tct ctg gaa gct gtc gtc cac gac atg gtg gag ctt 927 Thr Glu Gln His Ser Leu Glu Ala Val Val His Asp Met Val Glu Leu 270 275 280 gtg gct cag gca ctc agt agc atg gcc ctt gta cac cca gag cgg gca 975 Val Ala Gln Ala Leu Ser Ser Met Ala Leu Val His Pro Glu Arg Ala 285 290 295 300 ctg ctt cca gct gtg gtg aac tgt gat gac ctg aaa aca ggc gga tct 1023 Leu Leu Pro Ala Val Val Asn Cys Asp Asp Leu Lys Thr Gly Gly Ser 305 310 315 gag gca aca ggg cgc acc ttg gct cgg ttt ctc ggc aac acc tca ttt 1071 Glu Ala Thr Gly Arg Thr Leu Ala Arg Phe Leu Gly Asn Thr Ser Phe 320 325 330 cag ggc cga aca ggg gcc gtg tgg gtg aca ggc tcc tct cag gtg cat 1119 Gln Gly Arg Thr Gly Ala Val Trp Val Thr Gly Ser Ser Gln Val His 335 340 345 gtg tct cgg cat ttc aag gta tgg agc ctg cgc cgg gat ccg ctg ggt 1167 Val Ser Arg His Phe Lys Val Trp Ser Leu Arg Arg Asp Pro Leu Gly 350 355 360 gcc cca gcc tgg gca acc gtg ggc agc tgg cag gat gga cag ctg gac 1215 Ala Pro Ala Trp Ala Thr Val Gly Ser Trp Gln Asp Gly Gln Leu Asp 365 370 375 380 ttc cag cca ggg gca gcc gct ctc cga gtc cca tct ccg tct ggc acc 1263 Phe Gln Pro Gly Ala Ala Ala Leu Arg Val Pro Ser Pro Ser Gly Thr 385 390 395 cag gcc cga cca aag ctg cgt gtg gta acc ctg gtg gaa cac ccg ttt 1311 Gln Ala Arg Pro Lys Leu Arg Val Val Thr Leu Val Glu His Pro Phe 400 405 410 gtg ttc acc agg gaa tct gat gaa gac gga cag tgc cca gct ggg cag 1359 Val Phe Thr Arg Glu Ser Asp Glu Asp Gly Gln Cys Pro Ala Gly Gln 415 420 425 ctg tgt ctg gac cca ggc acc aat gac tca gcc agg ctg gat gcc ctc 1407 Leu Cys Leu Asp Pro Gly Thr Asn Asp Ser Ala Arg Leu Asp Ala Leu 430 435 440 ttt gct gca ctg gtg aat ggc tca gta cct cga acg ctg aga aga tgc 1455 Phe Ala Ala Leu Val Asn Gly Ser Val Pro Arg Thr Leu Arg Arg Cys 445 450 455 460 tgc tat ggc tac tgc atc gac ctg ctg gag cgg ctg gcc gag gac ctg 1503 Cys Tyr Gly Tyr Cys Ile Asp Leu Leu Glu Arg Leu Ala Glu Asp Leu 465 470 475 gcc ttt gac ttt gag ctc tat att gtg ggg gat ggc aag tac ggg gcc 1551 Ala Phe Asp Phe Glu Leu Tyr Ile Val Gly Asp Gly Lys Tyr Gly Ala 480 485 490 ctg cgt gat ggg cgc tgg acg ggc ctg gtg ggt gac ctg ctg gct ggc 1599 Leu Arg Asp Gly Arg Trp Thr Gly Leu Val Gly Asp Leu Leu Ala Gly 495 500 505 cgg gca cac atg gct gtg acc agc ttc agc atc aac tca gct cgc tct 1647 Arg Ala His Met Ala Val Thr Ser Phe Ser Ile Asn Ser Ala Arg Ser 510 515 520 cag gtg gtg gat ttc acc agc cct ttc ttc tcc acc agc ctg ggg att 1695 Gln Val Val Asp Phe Thr Ser Pro Phe Phe Ser Thr Ser Leu Gly Ile 525 530 535 540 atg gtg cgc acg aga gac acg gcc tcg ccc atc ggg gct ttc atg tgg 1743 Met Val Arg Thr Arg Asp Thr Ala Ser Pro Ile Gly Ala Phe Met Trp 545 550 555 ccc ctg cac tgg tcc atg tgg gtg ggc gtg ttt gct gct ctg cac ctc 1791 Pro Leu His Trp Ser Met Trp Val Gly Val Phe Ala Ala Leu His Leu 560 565 570 aca gcg ctc ttt ctc acc ctg tac gaa tgg cga agt ccc tac ggg ctc 1839 Thr Ala Leu Phe Leu Thr Leu Tyr Glu Trp Arg Ser Pro Tyr Gly Leu 575 580 585 acg ccg cgc ggc cgc aac cgt ggc act gtc ttc tct tac tcc tcc gcg 1887 Thr Pro Arg Gly Arg Asn Arg Gly Thr Val Phe Ser Tyr Ser Ser Ala 590 595 600 ctc aac ctg tgc tat gcc att ctc ttt gga cgc act gtc tcc agt aag 1935 Leu Asn Leu Cys Tyr Ala Ile Leu Phe Gly Arg Thr Val Ser Ser Lys 605 610 615 620 acg ccc aag tgc cct act gga cgc ttc ctc atg aac ctc tgg gca atc 1983 Thr Pro Lys Cys Pro Thr Gly Arg Phe Leu Met Asn Leu Trp Ala Ile 625 630 635 ttc tgc ctg ctg gtg ctt tcc agt tac acg gcc aac ctg gct gct gtc 2031 Phe Cys Leu Leu Val Leu Ser Ser Tyr Thr Ala Asn Leu Ala Ala Val 640 645 650 atg gtt ggg gac aaa acc ttt gag gag ctg tct gga atc cat gat ccc 2079 Met Val Gly Asp Lys Thr Phe Glu Glu Leu Ser Gly Ile His Asp Pro 655 660 665 aag ctg cac cac cct tcc caa ggc ttt cgc ttt ggc acc gta tgg gag 2127 Lys Leu His His Pro Ser Gln Gly Phe Arg Phe Gly Thr Val Trp Glu 670 675 680 agc agc gcg gag gcc tac atc aag gca agc ttc cct gag atg cac gca 2175 Ser Ser Ala Glu Ala Tyr Ile Lys Ala Ser Phe Pro Glu Met His Ala 685 690 695 700 cac atg cgt cgg cac agc gca ccc acc act cca cat ggg gtg gcc atg 2223 His Met Arg Arg His Ser Ala Pro Thr Thr Pro His Gly Val Ala Met 705 710 715 ctc acg agc gac ccg ccc aag ctc aac gcc ttc atc atg gat aaa tca 2271 Leu Thr Ser Asp Pro Pro Lys Leu Asn Ala Phe Ile Met Asp Lys Ser 720 725 730 cta ctg gac tat gag gtg tcc ata gat gcg gac tgc aag ctg ctc acc 2319 Leu Leu Asp Tyr Glu Val Ser Ile Asp Ala Asp Cys Lys Leu Leu Thr 735 740 745 gtt ggc aaa ccc ttt gct atc gag ggc tac ggc ata ggg cta ccc caa 2367 Val Gly Lys Pro Phe Ala Ile Glu Gly Tyr Gly Ile Gly Leu Pro Gln 750 755 760 aac tcg ccg ctc acc tcc aac ctg tcg gag ttc atc agt agg tac aag 2415 Asn Ser Pro Leu Thr Ser Asn Leu Ser Glu Phe Ile Ser Arg Tyr Lys 765 770 775 780 tct tca ggc ttc att gat ctg ctc cat gac aag tgg tac aag atg gtg 2463 Ser Ser Gly Phe Ile Asp Leu Leu His Asp Lys Trp Tyr Lys Met Val 785 790 795 cct tgc ggg aag cgg gtg ttc gcc gtg acg gag acg ctg cag atg ggg 2511 Pro Cys Gly Lys Arg Val Phe Ala Val Thr Glu Thr Leu Gln Met Gly 800 805 810 gtc tac cac ttc tca gga ttg ttt gtc ctg ctg tgc ctc ggg ctg ggc 2559 Val Tyr His Phe Ser Gly Leu Phe Val Leu Leu Cys Leu Gly Leu Gly 815 820 825 agc gcg ctt ctc acc tct ctg ggt gag cat gtc ttc tac cgc ctg gtg 2607 Ser Ala Leu Leu Thr Ser Leu Gly Glu His Val Phe Tyr Arg Leu Val 830 835 840 ctg ccg cgc atc cgc agg ggt aat aag ctg cag tat tgg ctt cac acg 2655 Leu Pro Arg Ile Arg Arg Gly Asn Lys Leu Gln Tyr Trp Leu His Thr 845 850 855 860 agc cag aag atc cac cga gcc ctc aat aca gga cca ccc gag ggg caa 2703 Ser Gln Lys Ile His Arg Ala Leu Asn Thr Gly Pro Pro Glu Gly Gln 865 870 875 cag gag agg gca gag cag gag cgc agc ggc ccc aag gac gag ctg cct 2751 Gln Glu Arg Ala Glu Gln Glu Arg Ser Gly Pro Lys Asp Glu Leu Pro 880 885 890 gcc acc gat ggt gca ggg cgc tgg agg cgg gtg cgc cgg gct gtg gaa 2799 Ala Thr Asp Gly Ala Gly Arg Trp Arg Arg Val Arg Arg Ala Val Glu 895 900 905 cgg gag cga cgc gtg cgt ttc ctg ctg gaa cct ggg gag gct ggc gga 2847 Arg Glu Arg Arg Val Arg Phe Leu Leu Glu Pro Gly Glu Ala Gly Gly 910 915 920 gac cgc ccg tgg ctc tgc tcc aac ggg ccc ggg ctg caa gcg gag ctg 2895 Asp Arg Pro Trp Leu Cys Ser Asn Gly Pro Gly Leu Gln Ala Glu Leu 925 930 935 940 cgg gag ctg gag ctg cgc att gag gct gca cgg gag cag ctg cgc agt 2943 Arg Glu Leu Glu Leu Arg Ile Glu Ala Ala Arg Glu Gln Leu Arg Ser 945 950 955 gcg ctg ttg cgg cgc ggg gag ctg cgg gcc ctg ctt ggg gat ggc acc 2991 Ala Leu Leu Arg Arg Gly Glu Leu Arg Ala Leu Leu Gly Asp Gly Thr 960 965 970 cgg ctc agg cca ctg cgc ctg ttg cat gcg gcg cct gct gag agc 3036 Arg Leu Arg Pro Leu Arg Leu Leu His Ala Ala Pro Ala Glu Ser 975 980 985 tgaggaacca caaggccgca ctgtccacga cagtttattc tatatacaaa cacgactctg 3096 tacactgcaa ttaaatagcg tggaacgtga aaaaaaa 3133 <210> SEQ ID NO 2 <211> LENGTH: 987 <212> TYPE: PRT <213> ORGANISM: Rattus sp. <400> SEQUENCE: 2 Met Glu Ser Val Arg Thr Leu Trp Leu Ser Val Ala Leu Ala Leu Ala 1 5 10 15 Val Gly Ser Arg Val Val Arg Gly His Pro Gln Pro Cys Arg Val Pro 20 25 30 Thr Arg Ala Gly Ala Ser Val Arg Leu Ala Ala Leu Leu Pro Arg Ala 35 40 45 Pro Ala Ala Arg Ala Arg Val Leu Ala Ala Leu Ala Thr Pro Ala Pro 50 55 60 Arg Leu Pro His Asn Leu Ser Leu Glu Leu Val Ala Val Ala Ser Pro 65 70 75 80 Thr Arg Asp Pro Ala Ser Leu Ala Arg Gly Leu Cys Gln Val Leu Ala 85 90 95 Pro Pro Gly Val Val Ala Ser Ile Ala Phe Pro Glu Ala Arg Pro Glu 100 105 110 Leu Arg Leu Leu Gln Phe Leu Ala Ala Ala Thr Glu Thr Pro Val Thr 115 120 125 Pro Phe His Leu Gln Leu Asp Trp Ala Ser Pro Leu Glu Thr Ile Leu 130 135 140 Asp Val Leu Val Ser Leu Val Arg Ala His Ala Trp Glu Asp Ile Ala 145 150 155 160 Leu Val Leu Cys Arg Val Arg Asp Pro Gly Ser Leu Val Thr Leu Trp 165 170 175 Thr Asn His Ala Ser Gln Ala Pro Lys Phe Val Leu Asp Leu Ser Arg 180 185 190 Leu Asp Ser Arg Asn Asp Ser Leu Arg Ala Gly Leu Ala Leu Leu Gly 195 200 205 Ala Leu Glu Gly Gly Gly Thr Pro Val Pro Ala Ala Val Leu Leu Gly 210 215 220 Cys Ser Thr Ala Arg Ala His Glu Val Leu Glu Ala Ala Pro Pro Gly 225 230 235 240 Pro Gln Trp Leu Leu Gly Thr Pro Leu Pro Ala Glu Ala Leu Pro Thr 245 250 255 Thr Gly Leu Pro Pro Gly Val Leu Ala Leu Gly Glu Thr Glu Gln His 260 265 270 Ser Leu Glu Ala Val Val His Asp Met Val Glu Leu Val Ala Gln Ala 275 280 285 Leu Ser Ser Met Ala Leu Val His Pro Glu Arg Ala Leu Leu Pro Ala 290 295 300 Val Val Asn Cys Asp Asp Leu Lys Thr Gly Gly Ser Glu Ala Thr Gly 305 310 315 320 Arg Thr Leu Ala Arg Phe Leu Gly Asn Thr Ser Phe Gln Gly Arg Thr 325 330 335 Gly Ala Val Trp Val Thr Gly Ser Ser Gln Val His Val Ser Arg His 340 345 350 Phe Lys Val Trp Ser Leu Arg Arg Asp Pro Leu Gly Ala Pro Ala Trp 355 360 365 Ala Thr Val Gly Ser Trp Gln Asp Gly Gln Leu Asp Phe Gln Pro Gly 370 375 380 Ala Ala Ala Leu Arg Val Pro Ser Pro Ser Gly Thr Gln Ala Arg Pro 385 390 395 400 Lys Leu Arg Val Val Thr Leu Val Glu His Pro Phe Val Phe Thr Arg 405 410 415 Glu Ser Asp Glu Asp Gly Gln Cys Pro Ala Gly Gln Leu Cys Leu Asp 420 425 430 Pro Gly Thr Asn Asp Ser Ala Arg Leu Asp Ala Leu Phe Ala Ala Leu 435 440 445 Val Asn Gly Ser Val Pro Arg Thr Leu Arg Arg Cys Cys Tyr Gly Tyr 450 455 460 Cys Ile Asp Leu Leu Glu Arg Leu Ala Glu Asp Leu Ala Phe Asp Phe 465 470 475 480 Glu Leu Tyr Ile Val Gly Asp Gly Lys Tyr Gly Ala Leu Arg Asp Gly 485 490 495 Arg Trp Thr Gly Leu Val Gly Asp Leu Leu Ala Gly Arg Ala His Met 500 505 510 Ala Val Thr Ser Phe Ser Ile Asn Ser Ala Arg Ser Gln Val Val Asp 515 520 525 Phe Thr Ser Pro Phe Phe Ser Thr Ser Leu Gly Ile Met Val Arg Thr 530 535 540 Arg Asp Thr Ala Ser Pro Ile Gly Ala Phe Met Trp Pro Leu His Trp 545 550 555 560 Ser Met Trp Val Gly Val Phe Ala Ala Leu His Leu Thr Ala Leu Phe 565 570 575 Leu Thr Leu Tyr Glu Trp Arg Ser Pro Tyr Gly Leu Thr Pro Arg Gly 580 585 590 Arg Asn Arg Gly Thr Val Phe Ser Tyr Ser Ser Ala Leu Asn Leu Cys 595 600 605 Tyr Ala Ile Leu Phe Gly Arg Thr Val Ser Ser Lys Thr Pro Lys Cys 610 615 620 Pro Thr Gly Arg Phe Leu Met Asn Leu Trp Ala Ile Phe Cys Leu Leu 625 630 635 640 Val Leu Ser Ser Tyr Thr Ala Asn Leu Ala Ala Val Met Val Gly Asp 645 650 655 Lys Thr Phe Glu Glu Leu Ser Gly Ile His Asp Pro Lys Leu His His 660 665 670 Pro Ser Gln Gly Phe Arg Phe Gly Thr Val Trp Glu Ser Ser Ala Glu 675 680 685 Ala Tyr Ile Lys Ala Ser Phe Pro Glu Met His Ala His Met Arg Arg 690 695 700 His Ser Ala Pro Thr Thr Pro His Gly Val Ala Met Leu Thr Ser Asp 705 710 715 720 Pro Pro Lys Leu Asn Ala Phe Ile Met Asp Lys Ser Leu Leu Asp Tyr 725 730 735 Glu Val Ser Ile Asp Ala Asp Cys Lys Leu Leu Thr Val Gly Lys Pro 740 745 750 Phe Ala Ile Glu Gly Tyr Gly Ile Gly Leu Pro Gln Asn Ser Pro Leu 755 760 765 Thr Ser Asn Leu Ser Glu Phe Ile Ser Arg Tyr Lys Ser Ser Gly Phe 770 775 780 Ile Asp Leu Leu His Asp Lys Trp Tyr Lys Met Val Pro Cys Gly Lys 785 790 795 800 Arg Val Phe Ala Val Thr Glu Thr Leu Gln Met Gly Val Tyr His Phe 805 810 815 Ser Gly Leu Phe Val Leu Leu Cys Leu Gly Leu Gly Ser Ala Leu Leu 820 825 830 Thr Ser Leu Gly Glu His Val Phe Tyr Arg Leu Val Leu Pro Arg Ile 835 840 845 Arg Arg Gly Asn Lys Leu Gln Tyr Trp Leu His Thr Ser Gln Lys Ile 850 855 860 His Arg Ala Leu Asn Thr Gly Pro Pro Glu Gly Gln Gln Glu Arg Ala 865 870 875 880 Glu Gln Glu Arg Ser Gly Pro Lys Asp Glu Leu Pro Ala Thr Asp Gly 885 890 895 Ala Gly Arg Trp Arg Arg Val Arg Arg Ala Val Glu Arg Glu Arg Arg 900 905 910 Val Arg Phe Leu Leu Glu Pro Gly Glu Ala Gly Gly Asp Arg Pro Trp 915 920 925 Leu Cys Ser Asn Gly Pro Gly Leu Gln Ala Glu Leu Arg Glu Leu Glu 930 935 940 Leu Arg Ile Glu Ala Ala Arg Glu Gln Leu Arg Ser Ala Leu Leu Arg 945 950 955 960 Arg Gly Glu Leu Arg Ala Leu Leu Gly Asp Gly Thr Arg Leu Arg Pro 965 970 975 Leu Arg Leu Leu His Ala Ala Pro Ala Glu Ser 980 985 <210> SEQ ID NO 3 <211> LENGTH: 3178 <212> TYPE: DNA <213> ORGANISM: Rattus sp. <220> FEATURE: <221> NAME/KEY: CDS <222> LOCATION: (76)...(3081) <400> SEQUENCE: 3 gcacgagggg acaagagcgg gtctggctgg ggtgtctcct tcccttcgca cagcacagtg 60 gtaacttctt tcggg atg gag agt gtg cgg acg ctg tgg ctc agc gtg gcc 111 Met Glu Ser Val Arg Thr Leu Trp Leu Ser Val Ala 1 5 10 ctg gcg ctg gcg gtg ggg tcc cga gtg gtg cgc ggt cac cct cag ccc 159 Leu Ala Leu Ala Val Gly Ser Arg Val Val Arg Gly His Pro Gln Pro 15 20 25 tgc cgg gtt ccc acg cgc gct ggg gcc tcc gtg cgc ctg gcg gcg ctc 207 Cys Arg Val Pro Thr Arg Ala Gly Ala Ser Val Arg Leu Ala Ala Leu 30 35 40 ctg ccc cgg gcg ccc gcc gcc cgc gcc cgc gtc cta gct gcc ctg gcc 255 Leu Pro Arg Ala Pro Ala Ala Arg Ala Arg Val Leu Ala Ala Leu Ala 45 50 55 60 acc cct gcg ccg cgg ctg ccg cac aac ctg agt ctg gaa ctg gtg gcc 303 Thr Pro Ala Pro Arg Leu Pro His Asn Leu Ser Leu Glu Leu Val Ala 65 70 75 gtc gcg tcc ccg acc cgg gac ccc gcg tcg cta gct cga ggt ctg tgc 351 Val Ala Ser Pro Thr Arg Asp Pro Ala Ser Leu Ala Arg Gly Leu Cys 80 85 90 cag gtt ctg gca ccg cct ggc gtg gtg gcc tct ata gcc ttt ccc gag 399 Gln Val Leu Ala Pro Pro Gly Val Val Ala Ser Ile Ala Phe Pro Glu 95 100 105 gcg cgg ccc gag ctg cgg cta ctg cag ttc ctg gca gcc gcc aca gag 447 Ala Arg Pro Glu Leu Arg Leu Leu Gln Phe Leu Ala Ala Ala Thr Glu 110 115 120 acc cca gtg gtg agc gtc ctg cgg agg gag gtg cgc acg gcc ctc gga 495 Thr Pro Val Val Ser Val Leu Arg Arg Glu Val Arg Thr Ala Leu Gly 125 130 135 140 gcc ccg act ccg ttc cat ctg cag ctg gac tgg gct agt ccc ctg gag 543 Ala Pro Thr Pro Phe His Leu Gln Leu Asp Trp Ala Ser Pro Leu Glu 145 150 155 acc ata ctg gat gtg ctg gtg tcc ctg gta cgg gca cat gcc tgg gag 591 Thr Ile Leu Asp Val Leu Val Ser Leu Val Arg Ala His Ala Trp Glu 160 165 170 gac att gct cta gta ctc tgc cgt gtc cgg gac cct ggc agc ctg gtg 639 Asp Ile Ala Leu Val Leu Cys Arg Val Arg Asp Pro Gly Ser Leu Val 175 180 185 aca ctc tgg act aac cat gct agc cag gct cca aag ttt gtg ctg gac 687 Thr Leu Trp Thr Asn His Ala Ser Gln Ala Pro Lys Phe Val Leu Asp 190 195 200 ctg agc cgg ctg gac agc agg aat gac agc ctt cgg gct gga ctg gcc 735 Leu Ser Arg Leu Asp Ser Arg Asn Asp Ser Leu Arg Ala Gly Leu Ala 205 210 215 220 ctg ttg ggg gcg ctg gaa gga ggg gga acc cca gtg cct gca gca gtc 783 Leu Leu Gly Ala Leu Glu Gly Gly Gly Thr Pro Val Pro Ala Ala Val 225 230 235 ctc cta ggc tgc agc act gcc cgt gca cat gag gtc cta gag gct gca 831 Leu Leu Gly Cys Ser Thr Ala Arg Ala His Glu Val Leu Glu Ala Ala 240 245 250 cca ccg ggt ccc cag tgg ttg ctg ggc aca cca ttg ccc gct gag gca 879 Pro Pro Gly Pro Gln Trp Leu Leu Gly Thr Pro Leu Pro Ala Glu Ala 255 260 265 ctg ccc acg act ggt ctg cca cct ggc gtg ctg gcg ctg ggg gaa acc 927 Leu Pro Thr Thr Gly Leu Pro Pro Gly Val Leu Ala Leu Gly Glu Thr 270 275 280 gaa caa cac tct ctg gaa gct gtc gtc cac gac atg gtg gag ctt gtg 975 Glu Gln His Ser Leu Glu Ala Val Val His Asp Met Val Glu Leu Val 285 290 295 300 gct cag gca ctc agt agc atg gcc ctt gta cac cca gag cgg gca ctg 1023 Ala Gln Ala Leu Ser Ser Met Ala Leu Val His Pro Glu Arg Ala Leu 305 310 315 ctt cca gct gtg gtg aac tgt gat gac ctg aaa aca ggc gga tct gag 1071 Leu Pro Ala Val Val Asn Cys Asp Asp Leu Lys Thr Gly Gly Ser Glu 320 325 330 gca aca ggg cgc acc ttg gct cgg ttt ctc ggc aac acc tca ttt cag 1119 Ala Thr Gly Arg Thr Leu Ala Arg Phe Leu Gly Asn Thr Ser Phe Gln 335 340 345 ggc cga aca ggg gcc gtg tgg gtg aca ggc tcc tct cag gtg cat gtg 1167 Gly Arg Thr Gly Ala Val Trp Val Thr Gly Ser Ser Gln Val His Val 350 355 360 tct cgg cat ttc aag gta tgg agc ctg cgc cgg gat ccg ctg ggt gcc 1215 Ser Arg His Phe Lys Val Trp Ser Leu Arg Arg Asp Pro Leu Gly Ala 365 370 375 380 cca gcc tgg gca acc gtg ggc agc tgg cag gat gga cag ctg gac ttc 1263 Pro Ala Trp Ala Thr Val Gly Ser Trp Gln Asp Gly Gln Leu Asp Phe 385 390 395 cag cca ggg gca gcc gct ctc cga gtc cca tct ccg tct ggc acc cag 1311 Gln Pro Gly Ala Ala Ala Leu Arg Val Pro Ser Pro Ser Gly Thr Gln 400 405 410 gcc cga cca aag ctg cgt gtg gta acc ctg gtg gaa cac ccg ttt gtg 1359 Ala Arg Pro Lys Leu Arg Val Val Thr Leu Val Glu His Pro Phe Val 415 420 425 ttc acc agg gaa tct gat gaa gac gga cag tgc cca gct ggg cag ctg 1407 Phe Thr Arg Glu Ser Asp Glu Asp Gly Gln Cys Pro Ala Gly Gln Leu 430 435 440 tgt ctg gac cca ggc acc aat gac tca gcc agg ctg gat gcc ctc ttt 1455 Cys Leu Asp Pro Gly Thr Asn Asp Ser Ala Arg Leu Asp Ala Leu Phe 445 450 455 460 gct gca ctg gtg aat ggc tca gta cct cga acg ctg aga aga tgc tgc 1503 Ala Ala Leu Val Asn Gly Ser Val Pro Arg Thr Leu Arg Arg Cys Cys 465 470 475 tat ggc tac tgc atc gac ctg ctg gag cgg ctg gcc gag gac ctg gcc 1551 Tyr Gly Tyr Cys Ile Asp Leu Leu Glu Arg Leu Ala Glu Asp Leu Ala 480 485 490 ttt gac ttt gag ctc tat att gtg ggg gat ggc aag tac ggg gcc ctg 1599 Phe Asp Phe Glu Leu Tyr Ile Val Gly Asp Gly Lys Tyr Gly Ala Leu 495 500 505 cgt gat ggg cgc tgg acg ggc ctg gtg ggt gac ctg ctg gct ggc cgg 1647 Arg Asp Gly Arg Trp Thr Gly Leu Val Gly Asp Leu Leu Ala Gly Arg 510 515 520 gca cac atg gct gtg acc agc ttc agc atc aac tca gct cgc tct cag 1695 Ala His Met Ala Val Thr Ser Phe Ser Ile Asn Ser Ala Arg Ser Gln 525 530 535 540 gtg gtg gat ttc acc agc cct ttc ttc tcc acc agc ctg ggg att atg 1743 Val Val Asp Phe Thr Ser Pro Phe Phe Ser Thr Ser Leu Gly Ile Met 545 550 555 gtg cgc acg aga gac acg gcc tcg ccc atc ggg gct ttc atg tgg ccc 1791 Val Arg Thr Arg Asp Thr Ala Ser Pro Ile Gly Ala Phe Met Trp Pro 560 565 570 ctg cac tgg tcc atg tgg gtg ggc gtg ttt gct gct ctg cac ctc aca 1839 Leu His Trp Ser Met Trp Val Gly Val Phe Ala Ala Leu His Leu Thr 575 580 585 gcg ctc ttt ctc acc ctg tac gaa tgg cga agt ccc tac ggg ctc acg 1887 Ala Leu Phe Leu Thr Leu Tyr Glu Trp Arg Ser Pro Tyr Gly Leu Thr 590 595 600 ccg cgc ggc cgc aac cgt ggc act gtc ttc tct tac tcc tcc gcg ctc 1935 Pro Arg Gly Arg Asn Arg Gly Thr Val Phe Ser Tyr Ser Ser Ala Leu 605 610 615 620 aac ctg tgc tat gcc att ctc ttt gga cgc act gtc tcc agt aag acg 1983 Asn Leu Cys Tyr Ala Ile Leu Phe Gly Arg Thr Val Ser Ser Lys Thr 625 630 635 ccc aag tgc cct act gga cgc ttc ctc atg aac ctc tgg gca atc ttc 2031 Pro Lys Cys Pro Thr Gly Arg Phe Leu Met Asn Leu Trp Ala Ile Phe 640 645 650 tgc ctg ctg gtg ctt tcc agt tac acg gcc aac ctg gct gct gtc atg 2079 Cys Leu Leu Val Leu Ser Ser Tyr Thr Ala Asn Leu Ala Ala Val Met 655 660 665 gtt ggg gac aaa acc ttt gag gag ctg tct gga atc cat gat ccc aag 2127 Val Gly Asp Lys Thr Phe Glu Glu Leu Ser Gly Ile His Asp Pro Lys 670 675 680 ctg cac cac cct tcc caa ggc ttt cgc ttt ggc acc gta tgg gag agc 2175 Leu His His Pro Ser Gln Gly Phe Arg Phe Gly Thr Val Trp Glu Ser 685 690 695 700 agc gcg gag gcc tac atc aag gca agc ttc cct gag atg cac gca cac 2223 Ser Ala Glu Ala Tyr Ile Lys Ala Ser Phe Pro Glu Met His Ala His 705 710 715 atg cgt cgg cac agc gca ccc acc act cca cat ggg gtg gcc atg ctc 2271 Met Arg Arg His Ser Ala Pro Thr Thr Pro His Gly Val Ala Met Leu 720 725 730 acg agc gac ccg ccc aag ctc aac gcc ttc atc atg gat aaa tca cta 2319 Thr Ser Asp Pro Pro Lys Leu Asn Ala Phe Ile Met Asp Lys Ser Leu 735 740 745 ctg gac tat gag gtg tcc ata gat gcg gac tgc aag ctg ctc acc gtt 2367 Leu Asp Tyr Glu Val Ser Ile Asp Ala Asp Cys Lys Leu Leu Thr Val 750 755 760 ggc aaa ccc ttt gct atc gag ggc tac ggc ata ggg cta ccc caa aac 2415 Gly Lys Pro Phe Ala Ile Glu Gly Tyr Gly Ile Gly Leu Pro Gln Asn 765 770 775 780 tcg ccg ctc acc tcc aac ctg tcg gag ttc atc agt agg tac aag tct 2463 Ser Pro Leu Thr Ser Asn Leu Ser Glu Phe Ile Ser Arg Tyr Lys Ser 785 790 795 tca ggc ttc att gat ctg ctc cat gac aag tgg tac aag atg gtg cct 2511 Ser Gly Phe Ile Asp Leu Leu His Asp Lys Trp Tyr Lys Met Val Pro 800 805 810 tgc ggg aag cgg gtg ttc gcc gtg acg gag acg ctg cag atg ggg gtc 2559 Cys Gly Lys Arg Val Phe Ala Val Thr Glu Thr Leu Gln Met Gly Val 815 820 825 tac cac ttc tca gga ttg ttt gtc ctg ctg tgc ctc ggg ctg ggc agc 2607 Tyr His Phe Ser Gly Leu Phe Val Leu Leu Cys Leu Gly Leu Gly Ser 830 835 840 gcg ctt ctc acc tct ctg ggt gag cat gtc ttc tac cgc ctg gtg ctg 2655 Ala Leu Leu Thr Ser Leu Gly Glu His Val Phe Tyr Arg Leu Val Leu 845 850 855 860 ccg cgc atc cgc agg ggt aat aag ctg cag tat tgg ctt cac acg agc 2703 Pro Arg Ile Arg Arg Gly Asn Lys Leu Gln Tyr Trp Leu His Thr Ser 865 870 875 cag aag atc cac cga gcc ctc aat aca gga cca ccc gag ggg caa cag 2751 Gln Lys Ile His Arg Ala Leu Asn Thr Gly Pro Pro Glu Gly Gln Gln 880 885 890 gag agg gca gag cag gag cgc agc ggc ccc aag gac gag ctg cct gcc 2799 Glu Arg Ala Glu Gln Glu Arg Ser Gly Pro Lys Asp Glu Leu Pro Ala 895 900 905 acc gat ggt gca ggg cgc tgg agg cgg gtg cgc cgg gct gtg gaa cgg 2847 Thr Asp Gly Ala Gly Arg Trp Arg Arg Val Arg Arg Ala Val Glu Arg 910 915 920 gag cga cgc gtg cgt ttc ctg ctg gaa cct ggg gag gct ggc gga gac 2895 Glu Arg Arg Val Arg Phe Leu Leu Glu Pro Gly Glu Ala Gly Gly Asp 925 930 935 940 cgc ccg tgg ctc tgc tcc aac ggg ccc ggg ctg caa gcg gag ctg cgg 2943 Arg Pro Trp Leu Cys Ser Asn Gly Pro Gly Leu Gln Ala Glu Leu Arg 945 950 955 gag ctg gag ctg cgc att gag gct gca cgg gag cag ctg cgc agt gcg 2991 Glu Leu Glu Leu Arg Ile Glu Ala Ala Arg Glu Gln Leu Arg Ser Ala 960 965 970 ctg ttg cgg cgc ggg gag ctg cgg gcc ctg ctt ggg gat ggc acc cgg 3039 Leu Leu Arg Arg Gly Glu Leu Arg Ala Leu Leu Gly Asp Gly Thr Arg 975 980 985 ctc agg cca ctg cgc ctg ttg cat gcg gcg cct gct gag agc 3081 Leu Arg Pro Leu Arg Leu Leu His Ala Ala Pro Ala Glu Ser 990 995 1000 tgaggaacca caaggccgca ctgtccacga cagtttattc tatatacaaa cacgactctg 3141 tacactgcaa ttaaatagcg tggaacgtga aaaaaaa 3178 <210> SEQ ID NO 4 <211> LENGTH: 1002 <212> TYPE: PRT <213> ORGANISM: Rattus sp. <400> SEQUENCE: 4 Met Glu Ser Val Arg Thr Leu Trp Leu Ser Val Ala Leu Ala Leu Ala 1 5 10 15 Val Gly Ser Arg Val Val Arg Gly His Pro Gln Pro Cys Arg Val Pro 20 25 30 Thr Arg Ala Gly Ala Ser Val Arg Leu Ala Ala Leu Leu Pro Arg Ala 35 40 45 Pro Ala Ala Arg Ala Arg Val Leu Ala Ala Leu Ala Thr Pro Ala Pro 50 55 60 Arg Leu Pro His Asn Leu Ser Leu Glu Leu Val Ala Val Ala Ser Pro 65 70 75 80 Thr Arg Asp Pro Ala Ser Leu Ala Arg Gly Leu Cys Gln Val Leu Ala 85 90 95 Pro Pro Gly Val Val Ala Ser Ile Ala Phe Pro Glu Ala Arg Pro Glu 100 105 110 Leu Arg Leu Leu Gln Phe Leu Ala Ala Ala Thr Glu Thr Pro Val Val 115 120 125 Ser Val Leu Arg Arg Glu Val Arg Thr Ala Leu Gly Ala Pro Thr Pro 130 135 140 Phe His Leu Gln Leu Asp Trp Ala Ser Pro Leu Glu Thr Ile Leu Asp 145 150 155 160 Val Leu Val Ser Leu Val Arg Ala His Ala Trp Glu Asp Ile Ala Leu 165 170 175 Val Leu Cys Arg Val Arg Asp Pro Gly Ser Leu Val Thr Leu Trp Thr 180 185 190 Asn His Ala Ser Gln Ala Pro Lys Phe Val Leu Asp Leu Ser Arg Leu 195 200 205 Asp Ser Arg Asn Asp Ser Leu Arg Ala Gly Leu Ala Leu Leu Gly Ala 210 215 220 Leu Glu Gly Gly Gly Thr Pro Val Pro Ala Ala Val Leu Leu Gly Cys 225 230 235 240 Ser Thr Ala Arg Ala His Glu Val Leu Glu Ala Ala Pro Pro Gly Pro 245 250 255 Gln Trp Leu Leu Gly Thr Pro Leu Pro Ala Glu Ala Leu Pro Thr Thr 260 265 270 Gly Leu Pro Pro Gly Val Leu Ala Leu Gly Glu Thr Glu Gln His Ser 275 280 285 Leu Glu Ala Val Val His Asp Met Val Glu Leu Val Ala Gln Ala Leu 290 295 300 Ser Ser Met Ala Leu Val His Pro Glu Arg Ala Leu Leu Pro Ala Val 305 310 315 320 Val Asn Cys Asp Asp Leu Lys Thr Gly Gly Ser Glu Ala Thr Gly Arg 325 330 335 Thr Leu Ala Arg Phe Leu Gly Asn Thr Ser Phe Gln Gly Arg Thr Gly 340 345 350 Ala Val Trp Val Thr Gly Ser Ser Gln Val His Val Ser Arg His Phe 355 360 365 Lys Val Trp Ser Leu Arg Arg Asp Pro Leu Gly Ala Pro Ala Trp Ala 370 375 380 Thr Val Gly Ser Trp Gln Asp Gly Gln Leu Asp Phe Gln Pro Gly Ala 385 390 395 400 Ala Ala Leu Arg Val Pro Ser Pro Ser Gly Thr Gln Ala Arg Pro Lys 405 410 415 Leu Arg Val Val Thr Leu Val Glu His Pro Phe Val Phe Thr Arg Glu 420 425 430 Ser Asp Glu Asp Gly Gln Cys Pro Ala Gly Gln Leu Cys Leu Asp Pro 435 440 445 Gly Thr Asn Asp Ser Ala Arg Leu Asp Ala Leu Phe Ala Ala Leu Val 450 455 460 Asn Gly Ser Val Pro Arg Thr Leu Arg Arg Cys Cys Tyr Gly Tyr Cys 465 470 475 480 Ile Asp Leu Leu Glu Arg Leu Ala Glu Asp Leu Ala Phe Asp Phe Glu 485 490 495 Leu Tyr Ile Val Gly Asp Gly Lys Tyr Gly Ala Leu Arg Asp Gly Arg 500 505 510 Trp Thr Gly Leu Val Gly Asp Leu Leu Ala Gly Arg Ala His Met Ala 515 520 525 Val Thr Ser Phe Ser Ile Asn Ser Ala Arg Ser Gln Val Val Asp Phe 530 535 540 Thr Ser Pro Phe Phe Ser Thr Ser Leu Gly Ile Met Val Arg Thr Arg 545 550 555 560 Asp Thr Ala Ser Pro Ile Gly Ala Phe Met Trp Pro Leu His Trp Ser 565 570 575 Met Trp Val Gly Val Phe Ala Ala Leu His Leu Thr Ala Leu Phe Leu 580 585 590 Thr Leu Tyr Glu Trp Arg Ser Pro Tyr Gly Leu Thr Pro Arg Gly Arg 595 600 605 Asn Arg Gly Thr Val Phe Ser Tyr Ser Ser Ala Leu Asn Leu Cys Tyr 610 615 620 Ala Ile Leu Phe Gly Arg Thr Val Ser Ser Lys Thr Pro Lys Cys Pro 625 630 635 640 Thr Gly Arg Phe Leu Met Asn Leu Trp Ala Ile Phe Cys Leu Leu Val 645 650 655 Leu Ser Ser Tyr Thr Ala Asn Leu Ala Ala Val Met Val Gly Asp Lys 660 665 670 Thr Phe Glu Glu Leu Ser Gly Ile His Asp Pro Lys Leu His His Pro 675 680 685 Ser Gln Gly Phe Arg Phe Gly Thr Val Trp Glu Ser Ser Ala Glu Ala 690 695 700 Tyr Ile Lys Ala Ser Phe Pro Glu Met His Ala His Met Arg Arg His 705 710 715 720 Ser Ala Pro Thr Thr Pro His Gly Val Ala Met Leu Thr Ser Asp Pro 725 730 735 Pro Lys Leu Asn Ala Phe Ile Met Asp Lys Ser Leu Leu Asp Tyr Glu 740 745 750 Val Ser Ile Asp Ala Asp Cys Lys Leu Leu Thr Val Gly Lys Pro Phe 755 760 765 Ala Ile Glu Gly Tyr Gly Ile Gly Leu Pro Gln Asn Ser Pro Leu Thr 770 775 780 Ser Asn Leu Ser Glu Phe Ile Ser Arg Tyr Lys Ser Ser Gly Phe Ile 785 790 795 800 Asp Leu Leu His Asp Lys Trp Tyr Lys Met Val Pro Cys Gly Lys Arg 805 810 815 Val Phe Ala Val Thr Glu Thr Leu Gln Met Gly Val Tyr His Phe Ser 820 825 830 Gly Leu Phe Val Leu Leu Cys Leu Gly Leu Gly Ser Ala Leu Leu Thr 835 840 845 Ser Leu Gly Glu His Val Phe Tyr Arg Leu Val Leu Pro Arg Ile Arg 850 855 860 Arg Gly Asn Lys Leu Gln Tyr Trp Leu His Thr Ser Gln Lys Ile His 865 870 875 880 Arg Ala Leu Asn Thr Gly Pro Pro Glu Gly Gln Gln Glu Arg Ala Glu 885 890 895 Gln Glu Arg Ser Gly Pro Lys Asp Glu Leu Pro Ala Thr Asp Gly Ala 900 905 910 Gly Arg Trp Arg Arg Val Arg Arg Ala Val Glu Arg Glu Arg Arg Val 915 920 925 Arg Phe Leu Leu Glu Pro Gly Glu Ala Gly Gly Asp Arg Pro Trp Leu 930 935 940 Cys Ser Asn Gly Pro Gly Leu Gln Ala Glu Leu Arg Glu Leu Glu Leu 945 950 955 960 Arg Ile Glu Ala Ala Arg Glu Gln Leu Arg Ser Ala Leu Leu Arg Arg 965 970 975 Gly Glu Leu Arg Ala Leu Leu Gly Asp Gly Thr Arg Leu Arg Pro Leu 980 985 990 Arg Leu Leu His Ala Ala Pro Ala Glu Ser 995 1000 <210> SEQ ID NO 5 <211> LENGTH: 3096 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <220> FEATURE: <221> NAME/KEY: CDS <222> LOCATION: (1)...(3033) <221> NAME/KEY: misc_feature <222> LOCATION: 2692, 2693, 2694, 2695, 2696, 2697, 2698, 2699, 2700, 2701, 2702, 2703, 2704, 2705, 2706, 2707, 2708, 2709, 2710, 2711, 2712, 2713, 2714, 2715, 2716, 2717, 2718, 2719, 2720, 2721, 2722, 2723, 2724, 2725, 2726, 2727, 2728, 2729, 2730 <223> OTHER INFORMATION: n = A,T,C or G <221> NAME/KEY: misc_feature <222> LOCATION: 2791, 2792, 2793, 2794, 2795, 2796, 2797, 2798, 2799, 2800, 2801, 2802, 2803, 2804, 2805, 2806, 2807, 2808, 2809, 2810, 2811, 2812, 2813, 2814, 2827, 2828, 2829, 2830, 2831, 2832, 2833, 2834, 2835, 2836, 2837, 2838, 2839, 2840, 2841 <223> OTHER INFORMATION: n = A,T,C or G <221> NAME/KEY: misc_feature <222> LOCATION: 2842, 2843, 2844, 2845, 2846, 2847, 3037, 3038, 3039 <223> OTHER INFORMATION: n = A,T,C or G <400> SEQUENCE: 5 atg gag ttt gtg cgg gcg ctg tgg ctg ggc ctg gcg ctg gcg ctg ggg 48 Met Glu Phe Val Arg Ala Leu Trp Leu Gly Leu Ala Leu Ala Leu Gly 1 5 10 15 ccg ggg tcc gcg ggg ggc cac cct cag ccg tgc ggc gtc ctg gcg cgc 96 Pro Gly Ser Ala Gly Gly His Pro Gln Pro Cys Gly Val Leu Ala Arg 20 25 30 ctc ggg ggc tcc gtg cgc ctg ggc gcc ctc ctg ccc cgc gcg cct ctc 144 Leu Gly Gly Ser Val Arg Leu Gly Ala Leu Leu Pro Arg Ala Pro Leu 35 40 45 gcc cgc gcc cgc gcc cgc gcc gcc ctg gcc cgg gcc gcc ctg gcg ccg 192 Ala Arg Ala Arg Ala Arg Ala Ala Leu Ala Arg Ala Ala Leu Ala Pro 50 55 60 cgg ctg ccg cac aac ctg agc ttg gag ctg gtg gtc gcc gcg ccc ccc 240 Arg Leu Pro His Asn Leu Ser Leu Glu Leu Val Val Ala Ala Pro Pro 65 70 75 80 gcc cgc gac ccc gcc tcg ctg acc cgc ggc ctg tgc cag gcg ctg gtg 288 Ala Arg Asp Pro Ala Ser Leu Thr Arg Gly Leu Cys Gln Ala Leu Val 85 90 95 cct ccg ggc gtg gcg gcc ctg ctc gcc ttt ccc gag gct cgg ccc gag 336 Pro Pro Gly Val Ala Ala Leu Leu Ala Phe Pro Glu Ala Arg Pro Glu 100 105 110 ctg ctg cag ctg cac ttc ctg gcg gcg gcc acc gag acc ccc gtg ctc 384 Leu Leu Gln Leu His Phe Leu Ala Ala Ala Thr Glu Thr Pro Val Leu 115 120 125 agc ctg ctg cgg cgg gag gcg cgc gcg ccc ctc gga gcc ccg aac cca 432 Ser Leu Leu Arg Arg Glu Ala Arg Ala Pro Leu Gly Ala Pro Asn Pro 130 135 140 ttc cac ctg cag ctg cac tgg gcc agc ccc ctg gag acg ctg ctg gat 480 Phe His Leu Gln Leu His Trp Ala Ser Pro Leu Glu Thr Leu Leu Asp 145 150 155 160 gtg ctg gtg gcg gtg ctg cag gcg cac gcc tgg gaa gac gtc ggc ctg 528 Val Leu Val Ala Val Leu Gln Ala His Ala Trp Glu Asp Val Gly Leu 165 170 175 gcc ctg tgc cgc act cag gac ccc ggc ggc ctg gtg gcc ctc tgg aca 576 Ala Leu Cys Arg Thr Gln Asp Pro Gly Gly Leu Val Ala Leu Trp Thr 180 185 190 agc cgg gct ggc cgg ccc cca cag ctg gtc ctg gac cta agc cgg cgg 624 Ser Arg Ala Gly Arg Pro Pro Gln Leu Val Leu Asp Leu Ser Arg Arg 195 200 205 gac acg gga gat gca gga ctg cgg gca cgc ctg gcc ccg atg gcg gcg 672 Asp Thr Gly Asp Ala Gly Leu Arg Ala Arg Leu Ala Pro Met Ala Ala 210 215 220 cca gtg ggg ggt gaa gca ccg gta ccc gcg gcg gtc ctc ctc ggc tgt 720 Pro Val Gly Gly Glu Ala Pro Val Pro Ala Ala Val Leu Leu Gly Cys 225 230 235 240 gac atc gcc cgt gcc cgt cgg gtg ctg gag gcc gta cct ccc ggc ccc 768 Asp Ile Ala Arg Ala Arg Arg Val Leu Glu Ala Val Pro Pro Gly Pro 245 250 255 cac tgg ctg ttg ggg aca cca ctg ccg ccc aag gcc ctg ccc acc gcg 816 His Trp Leu Leu Gly Thr Pro Leu Pro Pro Lys Ala Leu Pro Thr Ala 260 265 270 ggg ctg cca cca ggg ctg ctg gcg ctg ggc gag gtg gca cga ccc ccg 864 Gly Leu Pro Pro Gly Leu Leu Ala Leu Gly Glu Val Ala Arg Pro Pro 275 280 285 ctg gag gcc gcc atc cat gac att gtg caa ctg gtg gcc cgg gcg ctg 912 Leu Glu Ala Ala Ile His Asp Ile Val Gln Leu Val Ala Arg Ala Leu 290 295 300 ggc agt gcg gcc cag gtg cag ccg aag cga gcc ctc ctc ccc gcc ccg 960 Gly Ser Ala Ala Gln Val Gln Pro Lys Arg Ala Leu Leu Pro Ala Pro 305 310 315 320 gtc aac tgc ggg gac ctg cag ccg gcc ggg ccc gag tcc ccg ggg cgc 1008 Val Asn Cys Gly Asp Leu Gln Pro Ala Gly Pro Glu Ser Pro Gly Arg 325 330 335 ttc ttg gcc aac acg tcc ttc cag ggc cgc acg ggc ccc gtg tgg gtg 1056 Phe Leu Ala Asn Thr Ser Phe Gln Gly Arg Thr Gly Pro Val Trp Val 340 345 350 aca ggc agc tcc cag gta cac atg tct cgg cac ttt aag gtg tgg agc 1104 Thr Gly Ser Ser Gln Val His Met Ser Arg His Phe Lys Val Trp Ser 355 360 365 ctt cgc cgg gac cca cgg ggc gcc ccg gcc tgg gcc acg gtg ggc agc 1152 Leu Arg Arg Asp Pro Arg Gly Ala Pro Ala Trp Ala Thr Val Gly Ser 370 375 380 tgg cgg gac ggc cag ctg gac ttg gaa ccg gga ggt gcc tct gca cgg 1200 Trp Arg Asp Gly Gln Leu Asp Leu Glu Pro Gly Gly Ala Ser Ala Arg 385 390 395 400 ccc ccg ccc cca cag ggt gcc cag gtc tgg ccc aag ctg cgt gtg gta 1248 Pro Pro Pro Pro Gln Gly Ala Gln Val Trp Pro Lys Leu Arg Val Val 405 410 415 acg ctg ttg gaa cac cca ttt gtg ttt gcc cgt gat cca gac gaa gac 1296 Thr Leu Leu Glu His Pro Phe Val Phe Ala Arg Asp Pro Asp Glu Asp 420 425 430 ggg cag tgc cca gcg ggg cag ctg tgc ctg gac cct ggc acc aac gac 1344 Gly Gln Cys Pro Ala Gly Gln Leu Cys Leu Asp Pro Gly Thr Asn Asp 435 440 445 tcg gcc acc ctg gac gca ctg ttc gcc gcg ctg gcc aac ggc tca gcg 1392 Ser Ala Thr Leu Asp Ala Leu Phe Ala Ala Leu Ala Asn Gly Ser Ala 450 455 460 ccc cgt gcc ctg cgc aag tgc tgc tac ggc tac tgc att gac ctg ctg 1440 Pro Arg Ala Leu Arg Lys Cys Cys Tyr Gly Tyr Cys Ile Asp Leu Leu 465 470 475 480 gag cgg ctg gcg gag gac acg ccc ttc gac ttc gag ctg tac ctc gtg 1488 Glu Arg Leu Ala Glu Asp Thr Pro Phe Asp Phe Glu Leu Tyr Leu Val 485 490 495 ggt gac ggc aag tac ggc gcc ctg cgg gac ggc cgc tgg acc ggc ctg 1536 Gly Asp Gly Lys Tyr Gly Ala Leu Arg Asp Gly Arg Trp Thr Gly Leu 500 505 510 gtc ggg gac ctg ctg gcc ggc cgg gcc cac atg gcg gtc acc agc ttc 1584 Val Gly Asp Leu Leu Ala Gly Arg Ala His Met Ala Val Thr Ser Phe 515 520 525 agt atc aac tcc gcc cgc tca cag gtg gtg gac ttc acc agc ccc ttc 1632 Ser Ile Asn Ser Ala Arg Ser Gln Val Val Asp Phe Thr Ser Pro Phe 530 535 540 ttc tcc acc agc ctg ggc atc atg gtg cgg gca cgg gac acg gcc tca 1680 Phe Ser Thr Ser Leu Gly Ile Met Val Arg Ala Arg Asp Thr Ala Ser 545 550 555 560 ccc atc ggt gcc ttt atg tgg ccc ctg cac tgg tcc acg tgg ctg ggc 1728 Pro Ile Gly Ala Phe Met Trp Pro Leu His Trp Ser Thr Trp Leu Gly 565 570 575 gtc ttt gcg gcc ctg cac ctc acc gcg ctc ttc ctc acc gtg tac gag 1776 Val Phe Ala Ala Leu His Leu Thr Ala Leu Phe Leu Thr Val Tyr Glu 580 585 590 tgg cgt agc ccc tac ggc ctc acg cca cgt ggc cgc aac cgc agc acc 1824 Trp Arg Ser Pro Tyr Gly Leu Thr Pro Arg Gly Arg Asn Arg Ser Thr 595 600 605 gtc ttc tcc tac tcc tca gcc ctc aac ctg tgc tac gcc atc ctc ttc 1872 Val Phe Ser Tyr Ser Ser Ala Leu Asn Leu Cys Tyr Ala Ile Leu Phe 610 615 620 aga cgc acc gtg tcc agc aag acg ccc aag tgc ccc acg ggc cgc ctg 1920 Arg Arg Thr Val Ser Ser Lys Thr Pro Lys Cys Pro Thr Gly Arg Leu 625 630 635 640 ctc atg aac ctc tgg gcc atc ttc tgc ctg ctg gtg ctg tcc agc tac 1968 Leu Met Asn Leu Trp Ala Ile Phe Cys Leu Leu Val Leu Ser Ser Tyr 645 650 655 acg gcc aac ctg gct gcc gtc atg gtc ggg gac aag acc ttc gag gag 2016 Thr Ala Asn Leu Ala Ala Val Met Val Gly Asp Lys Thr Phe Glu Glu 660 665 670 ctg tcg ggg atc cac gac ccc aag ctg cac cac ccg gcg cag ggc ttc 2064 Leu Ser Gly Ile His Asp Pro Lys Leu His His Pro Ala Gln Gly Phe 675 680 685 cgc ttc ggc acc gtg tgg gag agc agc gcc gag gcg tac atc aag aag 2112 Arg Phe Gly Thr Val Trp Glu Ser Ser Ala Glu Ala Tyr Ile Lys Lys 690 695 700 agc ttc ccc gac atg cac gca cac atg cgg cgc cac agc gcg ccc acc 2160 Ser Phe Pro Asp Met His Ala His Met Arg Arg His Ser Ala Pro Thr 705 710 715 720 acg ccc cgc ggc gtc gcc atg ctc acg agc gac ccc ccc aag ctc aac 2208 Thr Pro Arg Gly Val Ala Met Leu Thr Ser Asp Pro Pro Lys Leu Asn 725 730 735 gcc ttc atc atg gac aag tcg ctc ctg gac tac gag gtc tcc atc gac 2256 Ala Phe Ile Met Asp Lys Ser Leu Leu Asp Tyr Glu Val Ser Ile Asp 740 745 750 gcc gac tgc aaa ctg ctg acc gtg gga aag ccc ttc gcc att gag ggc 2304 Ala Asp Cys Lys Leu Leu Thr Val Gly Lys Pro Phe Ala Ile Glu Gly 755 760 765 tat ggg atc gga ctg ccc cag aac tcg ccg ctc acc tcc aac ctg tcc 2352 Tyr Gly Ile Gly Leu Pro Gln Asn Ser Pro Leu Thr Ser Asn Leu Ser 770 775 780 gag ttc atc agc cgc tac aag tcc tcc ggc ttc atc gac ctg ctc cac 2400 Glu Phe Ile Ser Arg Tyr Lys Ser Ser Gly Phe Ile Asp Leu Leu His 785 790 795 800 gac aag tgg tac aag atg gtg cct tgc ggc aag cgg gtc ttt gcg gtt 2448 Asp Lys Trp Tyr Lys Met Val Pro Cys Gly Lys Arg Val Phe Ala Val 805 810 815 aca gag acc ctg cag atg agc atc tac cac ttc gcg ggc ctc ttc gtg 2496 Thr Glu Thr Leu Gln Met Ser Ile Tyr His Phe Ala Gly Leu Phe Val 820 825 830 ttg ctg tgc ctg ggc ctg ggc agc gct ctg ctc agc tcg ctg ggc gag 2544 Leu Leu Cys Leu Gly Leu Gly Ser Ala Leu Leu Ser Ser Leu Gly Glu 835 840 845 cac gcc ttc ttc cgc ctg gcg ctg ccg cgc atc cgc aag ggg agc agg 2592 His Ala Phe Phe Arg Leu Ala Leu Pro Arg Ile Arg Lys Gly Ser Arg 850 855 860 ctg cag tac tgg ctg cac acc agc cag aaa atc cac cgc gcc ctc aac 2640 Leu Gln Tyr Trp Leu His Thr Ser Gln Lys Ile His Arg Ala Leu Asn 865 870 875 880 acg gag cca cca gag ggg tcg aag gag gag acg gca gag gcg gag ccc 2688 Thr Glu Pro Pro Glu Gly Ser Lys Glu Glu Thr Ala Glu Ala Glu Pro 885 890 895 agg nnn nnn nnn nnn nnn nnn nnn nnn nnn nnn nnn nnn nnn tgg aaa 2736 Arg Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Trp Lys 900 905 910 cgg gcg cgc cgg gcc gtg gac aag gag cgc cgc gtg cgc ttc ctg ctg 2784 Arg Ala Arg Arg Ala Val Asp Lys Glu Arg Arg Val Arg Phe Leu Leu 915 920 925 gag ccc nnn nnn nnn nnn nnn nnn nnn nnn tgg ctg tgc tcc nnn nnn 2832 Glu Pro Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Trp Leu Cys Ser Xaa Xaa 930 935 940 nnn nnn nnn nnn nnn gag ctg cag gag ctg gag cgc cgc atc gaa gtc 2880 Xaa Xaa Xaa Xaa Xaa Glu Leu Gln Glu Leu Glu Arg Arg Ile Glu Val 945 950 955 960 gcg cgt gag cgg ctc cgc cag gcc ctg gtg cgg cgc ggc cag ctc ctg 2928 Ala Arg Glu Arg Leu Arg Gln Ala Leu Val Arg Arg Gly Gln Leu Leu 965 970 975 gca cag ctc ggg gac agc gca cgt cac cgg cct cgg cgc ttg ctt cag 2976 Ala Gln Leu Gly Asp Ser Ala Arg His Arg Pro Arg Arg Leu Leu Gln 980 985 990 gcc aga gcg gcc ccc gcg gag gcc cca cca cac tct ggc cga ccg ggg 3024 Ala Arg Ala Ala Pro Ala Glu Ala Pro Pro His Ser Gly Arg Pro Gly 995 1000 1005 agc cag gaa tgannngaca gtttattcta tatacaaaca caattttgta 3073 Ser Gln Glu 1010 cactgcaatt aaatagaatg gaa 3096 <210> SEQ ID NO 6 <211> LENGTH: 1011 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <220> FEATURE: <221> NAME/KEY: VARIANT <222> LOCATION: 898, 899, 900, 901, 902, 903, 904, 905, 906, 907, 908, 909, 910, 931, 932, 933, 934, 935, 936, 937, 938, 943, 944, 945, 946, 947, 948, 949 <223> OTHER INFORMATION: Xaa = Any Amino Acid <400> SEQUENCE: 6 Met Glu Phe Val Arg Ala Leu Trp Leu Gly Leu Ala Leu Ala Leu Gly 1 5 10 15 Pro Gly Ser Ala Gly Gly His Pro Gln Pro Cys Gly Val Leu Ala Arg 20 25 30 Leu Gly Gly Ser Val Arg Leu Gly Ala Leu Leu Pro Arg Ala Pro Leu 35 40 45 Ala Arg Ala Arg Ala Arg Ala Ala Leu Ala Arg Ala Ala Leu Ala Pro 50 55 60 Arg Leu Pro His Asn Leu Ser Leu Glu Leu Val Val Ala Ala Pro Pro 65 70 75 80 Ala Arg Asp Pro Ala Ser Leu Thr Arg Gly Leu Cys Gln Ala Leu Val 85 90 95 Pro Pro Gly Val Ala Ala Leu Leu Ala Phe Pro Glu Ala Arg Pro Glu 100 105 110 Leu Leu Gln Leu His Phe Leu Ala Ala Ala Thr Glu Thr Pro Val Leu 115 120 125 Ser Leu Leu Arg Arg Glu Ala Arg Ala Pro Leu Gly Ala Pro Asn Pro 130 135 140 Phe His Leu Gln Leu His Trp Ala Ser Pro Leu Glu Thr Leu Leu Asp 145 150 155 160 Val Leu Val Ala Val Leu Gln Ala His Ala Trp Glu Asp Val Gly Leu 165 170 175 Ala Leu Cys Arg Thr Gln Asp Pro Gly Gly Leu Val Ala Leu Trp Thr 180 185 190 Ser Arg Ala Gly Arg Pro Pro Gln Leu Val Leu Asp Leu Ser Arg Arg 195 200 205 Asp Thr Gly Asp Ala Gly Leu Arg Ala Arg Leu Ala Pro Met Ala Ala 210 215 220 Pro Val Gly Gly Glu Ala Pro Val Pro Ala Ala Val Leu Leu Gly Cys 225 230 235 240 Asp Ile Ala Arg Ala Arg Arg Val Leu Glu Ala Val Pro Pro Gly Pro 245 250 255 His Trp Leu Leu Gly Thr Pro Leu Pro Pro Lys Ala Leu Pro Thr Ala 260 265 270 Gly Leu Pro Pro Gly Leu Leu Ala Leu Gly Glu Val Ala Arg Pro Pro 275 280 285 Leu Glu Ala Ala Ile His Asp Ile Val Gln Leu Val Ala Arg Ala Leu 290 295 300 Gly Ser Ala Ala Gln Val Gln Pro Lys Arg Ala Leu Leu Pro Ala Pro 305 310 315 320 Val Asn Cys Gly Asp Leu Gln Pro Ala Gly Pro Glu Ser Pro Gly Arg 325 330 335 Phe Leu Ala Asn Thr Ser Phe Gln Gly Arg Thr Gly Pro Val Trp Val 340 345 350 Thr Gly Ser Ser Gln Val His Met Ser Arg His Phe Lys Val Trp Ser 355 360 365 Leu Arg Arg Asp Pro Arg Gly Ala Pro Ala Trp Ala Thr Val Gly Ser 370 375 380 Trp Arg Asp Gly Gln Leu Asp Leu Glu Pro Gly Gly Ala Ser Ala Arg 385 390 395 400 Pro Pro Pro Pro Gln Gly Ala Gln Val Trp Pro Lys Leu Arg Val Val 405 410 415 Thr Leu Leu Glu His Pro Phe Val Phe Ala Arg Asp Pro Asp Glu Asp 420 425 430 Gly Gln Cys Pro Ala Gly Gln Leu Cys Leu Asp Pro Gly Thr Asn Asp 435 440 445 Ser Ala Thr Leu Asp Ala Leu Phe Ala Ala Leu Ala Asn Gly Ser Ala 450 455 460 Pro Arg Ala Leu Arg Lys Cys Cys Tyr Gly Tyr Cys Ile Asp Leu Leu 465 470 475 480 Glu Arg Leu Ala Glu Asp Thr Pro Phe Asp Phe Glu Leu Tyr Leu Val 485 490 495 Gly Asp Gly Lys Tyr Gly Ala Leu Arg Asp Gly Arg Trp Thr Gly Leu 500 505 510 Val Gly Asp Leu Leu Ala Gly Arg Ala His Met Ala Val Thr Ser Phe 515 520 525 Ser Ile Asn Ser Ala Arg Ser Gln Val Val Asp Phe Thr Ser Pro Phe 530 535 540 Phe Ser Thr Ser Leu Gly Ile Met Val Arg Ala Arg Asp Thr Ala Ser 545 550 555 560 Pro Ile Gly Ala Phe Met Trp Pro Leu His Trp Ser Thr Trp Leu Gly 565 570 575 Val Phe Ala Ala Leu His Leu Thr Ala Leu Phe Leu Thr Val Tyr Glu 580 585 590 Trp Arg Ser Pro Tyr Gly Leu Thr Pro Arg Gly Arg Asn Arg Ser Thr 595 600 605 Val Phe Ser Tyr Ser Ser Ala Leu Asn Leu Cys Tyr Ala Ile Leu Phe 610 615 620 Arg Arg Thr Val Ser Ser Lys Thr Pro Lys Cys Pro Thr Gly Arg Leu 625 630 635 640 Leu Met Asn Leu Trp Ala Ile Phe Cys Leu Leu Val Leu Ser Ser Tyr 645 650 655 Thr Ala Asn Leu Ala Ala Val Met Val Gly Asp Lys Thr Phe Glu Glu 660 665 670 Leu Ser Gly Ile His Asp Pro Lys Leu His His Pro Ala Gln Gly Phe 675 680 685 Arg Phe Gly Thr Val Trp Glu Ser Ser Ala Glu Ala Tyr Ile Lys Lys 690 695 700 Ser Phe Pro Asp Met His Ala His Met Arg Arg His Ser Ala Pro Thr 705 710 715 720 Thr Pro Arg Gly Val Ala Met Leu Thr Ser Asp Pro Pro Lys Leu Asn 725 730 735 Ala Phe Ile Met Asp Lys Ser Leu Leu Asp Tyr Glu Val Ser Ile Asp 740 745 750 Ala Asp Cys Lys Leu Leu Thr Val Gly Lys Pro Phe Ala Ile Glu Gly 755 760 765 Tyr Gly Ile Gly Leu Pro Gln Asn Ser Pro Leu Thr Ser Asn Leu Ser 770 775 780 Glu Phe Ile Ser Arg Tyr Lys Ser Ser Gly Phe Ile Asp Leu Leu His 785 790 795 800 Asp Lys Trp Tyr Lys Met Val Pro Cys Gly Lys Arg Val Phe Ala Val 805 810 815 Thr Glu Thr Leu Gln Met Ser Ile Tyr His Phe Ala Gly Leu Phe Val 820 825 830 Leu Leu Cys Leu Gly Leu Gly Ser Ala Leu Leu Ser Ser Leu Gly Glu 835 840 845 His Ala Phe Phe Arg Leu Ala Leu Pro Arg Ile Arg Lys Gly Ser Arg 850 855 860 Leu Gln Tyr Trp Leu His Thr Ser Gln Lys Ile His Arg Ala Leu Asn 865 870 875 880 Thr Glu Pro Pro Glu Gly Ser Lys Glu Glu Thr Ala Glu Ala Glu Pro 885 890 895 Arg Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Trp Lys 900 905 910 Arg Ala Arg Arg Ala Val Asp Lys Glu Arg Arg Val Arg Phe Leu Leu 915 920 925 Glu Pro Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Trp Leu Cys Ser Xaa Xaa 930 935 940 Xaa Xaa Xaa Xaa Xaa Glu Leu Gln Glu Leu Glu Arg Arg Ile Glu Val 945 950 955 960 Ala Arg Glu Arg Leu Arg Gln Ala Leu Val Arg Arg Gly Gln Leu Leu 965 970 975 Ala Gln Leu Gly Asp Ser Ala Arg His Arg Pro Arg Arg Leu Leu Gln 980 985 990 Ala Arg Ala Ala Pro Ala Glu Ala Pro Pro His Ser Gly Arg Pro Gly 995 1000 1005 Ser Gln Glu 1010 <210> SEQ ID NO 7 <211> LENGTH: 3166 <212> TYPE: DNA <213> ORGANISM: Mus musculus <220> FEATURE: <221> NAME/KEY: CDS <222> LOCATION: (63)...(3077) <400> SEQUENCE: 7 agagcggggc tggctgggat gtctccttac cctcgcacag cacagtggta acttccatcg 60 gg atg gag tgt gtg cag acg ctg tgg ctc agc ctg gcc ctg gcg ctg 107 Met Glu Cys Val Gln Thr Leu Trp Leu Ser Leu Ala Leu Ala Leu 1 5 10 15 gcg cga ggg tcc tgg gtg gtg cgc ggt cac cct cag ccc tgc ggg gtt 155 Ala Arg Gly Ser Trp Val Val Arg Gly His Pro Gln Pro Cys Gly Val 20 25 30 ccc acg cgc gcc ggg gcc tcc gtg cgc ctg gct gcg ctc cta ccc cgg 203 Pro Thr Arg Ala Gly Ala Ser Val Arg Leu Ala Ala Leu Leu Pro Arg 35 40 45 gcg ccc gcc gcc cgc gcc cgc gtc cta gcc gcc ctg gcc acc cct tcc 251 Ala Pro Ala Ala Arg Ala Arg Val Leu Ala Ala Leu Ala Thr Pro Ser 50 55 60 ccg cgg ctg ccg cac aac ctg agt ctg gag cta gtg gcc gtc gcg tcc 299 Pro Arg Leu Pro His Asn Leu Ser Leu Glu Leu Val Ala Val Ala Ser 65 70 75 cca acc cgg gac ccc gcg tcg ctg gcc cga ggt ctg tgc cag gtt ctg 347 Pro Thr Arg Asp Pro Ala Ser Leu Ala Arg Gly Leu Cys Gln Val Leu 80 85 90 95 gca ccg ccc ggc gtg gtg gcc tct ata acc ttt ccc gag gcg cgg cct 395 Ala Pro Pro Gly Val Val Ala Ser Ile Thr Phe Pro Glu Ala Arg Pro 100 105 110 gag cta cgg cta ttg cag ttc ctg gca gct gcc aca gag acc ccg gtg 443 Glu Leu Arg Leu Leu Gln Phe Leu Ala Ala Ala Thr Glu Thr Pro Val 115 120 125 ctg agc gtc cta cgg agg gag gtg cgc gcg ccc ctc gga gct cgg cgg 491 Leu Ser Val Leu Arg Arg Glu Val Arg Ala Pro Leu Gly Ala Arg Arg 130 135 140 acc ccg ttc cac ctg cag ctg gac tgg gct agt ccc ctg gag acc atc 539 Thr Pro Phe His Leu Gln Leu Asp Trp Ala Ser Pro Leu Glu Thr Ile 145 150 155 ctg gat gtg ctg gtg tcc ctg gta cgg gca cac gcc tgg gag gac att 587 Leu Asp Val Leu Val Ser Leu Val Arg Ala His Ala Trp Glu Asp Ile 160 165 170 175 gct cta gtg ctc tgt cgt gtc cgg gac ccc agt ggc ctg gtg aca ctc 635 Ala Leu Val Leu Cys Arg Val Arg Asp Pro Ser Gly Leu Val Thr Leu 180 185 190 tgg acc agc cgt gct agc cag gct cca aag ttt gtg ctg gac ctg agc 683 Trp Thr Ser Arg Ala Ser Gln Ala Pro Lys Phe Val Leu Asp Leu Ser 195 200 205 cag ttg gac agc ggg aat gac agc ctt cgg gct aca ctg gcc ctg ctg 731 Gln Leu Asp Ser Gly Asn Asp Ser Leu Arg Ala Thr Leu Ala Leu Leu 210 215 220 ggg acg ctg gaa gga ggg gga acc ccc gtg tct gca gcc gtc ctc ctg 779 Gly Thr Leu Glu Gly Gly Gly Thr Pro Val Ser Ala Ala Val Leu Leu 225 230 235 ggc tgc agc act gcc cat gca cat gag gtc cta gag gca gca cca ccg 827 Gly Cys Ser Thr Ala His Ala His Glu Val Leu Glu Ala Ala Pro Pro 240 245 250 255 ggt ccc cag tgg ctg ctg ggc aca cca ctg ccc gcc gag gca ctg ccc 875 Gly Pro Gln Trp Leu Leu Gly Thr Pro Leu Pro Ala Glu Ala Leu Pro 260 265 270 aaa acc ggt ctg ccc cct ggg gtg ttg gtg ctg ggg gaa acc ggg cag 923 Lys Thr Gly Leu Pro Pro Gly Val Leu Val Leu Gly Glu Thr Gly Gln 275 280 285 cct tcc ctg gaa gct gcc gtc cac gac atg gtg gag ctt gtg gct cgg 971 Pro Ser Leu Glu Ala Ala Val His Asp Met Val Glu Leu Val Ala Arg 290 295 300 gca ctc agc agc atg gcc ctc atg cac cca gag cgg gcc ctg ctt cca 1019 Ala Leu Ser Ser Met Ala Leu Met His Pro Glu Arg Ala Leu Leu Pro 305 310 315 gcg gca gta aat tgt gag gac ctg aaa acg ggc ggc tct gag tca aca 1067 Ala Ala Val Asn Cys Glu Asp Leu Lys Thr Gly Gly Ser Glu Ser Thr 320 325 330 335 gca cgc acc ttg gct agg tgg ttt ctg agc aac acc tca ttt cag ggc 1115 Ala Arg Thr Leu Ala Arg Trp Phe Leu Ser Asn Thr Ser Phe Gln Gly 340 345 350 cgc aca ggg gct gtg tgg gtg gca ggc tcc tct cag gtg cat gtg tct 1163 Arg Thr Gly Ala Val Trp Val Ala Gly Ser Ser Gln Val His Val Ser 355 360 365 cgg cat ttc aag gta tgg agc tta cgc agg gac ccg ctg ggt gcc cca 1211 Arg His Phe Lys Val Trp Ser Leu Arg Arg Asp Pro Leu Gly Ala Pro 370 375 380 gcc tgg gca aca gta ggc agc tgg cag gat gga cag ctg gac ttc cag 1259 Ala Trp Ala Thr Val Gly Ser Trp Gln Asp Gly Gln Leu Asp Phe Gln 385 390 395 cca ggg gcg gct gct ctc cga gtt cca tct cca tct ggc acc cag gcc 1307 Pro Gly Ala Ala Ala Leu Arg Val Pro Ser Pro Ser Gly Thr Gln Ala 400 405 410 415 cgg cca aag ctg cga gtg gta act ctg gtg gaa cac cca ttt gtg ttc 1355 Arg Pro Lys Leu Arg Val Val Thr Leu Val Glu His Pro Phe Val Phe 420 425 430 acc agg gaa tct gat gaa gat ggg cag tgc ccg gct ggg cag ctg tgt 1403 Thr Arg Glu Ser Asp Glu Asp Gly Gln Cys Pro Ala Gly Gln Leu Cys 435 440 445 cta gac cca ggc acc aat gac tcg gcc agg ctg gat gcg ctc ttc act 1451 Leu Asp Pro Gly Thr Asn Asp Ser Ala Arg Leu Asp Ala Leu Phe Thr 450 455 460 gca ttg gag aat ggc tcc gtg cct cgc acc ctg aga aga tgc tgt tat 1499 Ala Leu Glu Asn Gly Ser Val Pro Arg Thr Leu Arg Arg Cys Cys Tyr 465 470 475 ggc tac tgc att gac ctg ctg gag cgg ctg gcc gag gac ctg gcc ttt 1547 Gly Tyr Cys Ile Asp Leu Leu Glu Arg Leu Ala Glu Asp Leu Ala Phe 480 485 490 495 gac ttt gag ctc tat att gtg ggg gat ggc aag tac ggg gcc ctg cgt 1595 Asp Phe Glu Leu Tyr Ile Val Gly Asp Gly Lys Tyr Gly Ala Leu Arg 500 505 510 gat gga cgc tgg aca ggc ctg gtg ggt gac ctg ctg gcc ggt cgg gca 1643 Asp Gly Arg Trp Thr Gly Leu Val Gly Asp Leu Leu Ala Gly Arg Ala 515 520 525 cac atg gcc gtg acc agc ttc agt atc aac tcg gct cgc tcc cag gtg 1691 His Met Ala Val Thr Ser Phe Ser Ile Asn Ser Ala Arg Ser Gln Val 530 535 540 gtg gat ttc acc agc cct ttc ttc tcc acc agc ctg ggc atc atg gtg 1739 Val Asp Phe Thr Ser Pro Phe Phe Ser Thr Ser Leu Gly Ile Met Val 545 550 555 cgc acg cga gat aca gcc tca ccc att ggg gct ttc atg tgg ccc ctg 1787 Arg Thr Arg Asp Thr Ala Ser Pro Ile Gly Ala Phe Met Trp Pro Leu 560 565 570 575 cac tgg tcc atg tgg gtg ggt gtg ttt gct gcc ctg cac ctc aca gcg 1835 His Trp Ser Met Trp Val Gly Val Phe Ala Ala Leu His Leu Thr Ala 580 585 590 ctc ttc ctt act ctg tac gaa tgg cgg agt ccc tac ggg ctc acg cca 1883 Leu Phe Leu Thr Leu Tyr Glu Trp Arg Ser Pro Tyr Gly Leu Thr Pro 595 600 605 cgc ggc cgc aac cgt ggt acc gtc ttc tcc tac tcc tcc gct ctc aac 1931 Arg Gly Arg Asn Arg Gly Thr Val Phe Ser Tyr Ser Ser Ala Leu Asn 610 615 620 ctc tgc tac gcc att ctc ttc gga cgc act gtc tcc agt aag aca ccc 1979 Leu Cys Tyr Ala Ile Leu Phe Gly Arg Thr Val Ser Ser Lys Thr Pro 625 630 635 aag tgt cct acc gga cgc ttc ctc atg aat ctc tgg gca atc ttc tgc 2027 Lys Cys Pro Thr Gly Arg Phe Leu Met Asn Leu Trp Ala Ile Phe Cys 640 645 650 655 ctg ctg gtg cta tcc agt tac aca gcc aac ctg gca gct gtc atg gtc 2075 Leu Leu Val Leu Ser Ser Tyr Thr Ala Asn Leu Ala Ala Val Met Val 660 665 670 ggg gac aag aca ttt gag gag ctg tct gga atc cat gat ccc aag ctg 2123 Gly Asp Lys Thr Phe Glu Glu Leu Ser Gly Ile His Asp Pro Lys Leu 675 680 685 cac cac cct tcc caa ggc ttc cgc ttt ggc acc gtg tgg gag agc agc 2171 His His Pro Ser Gln Gly Phe Arg Phe Gly Thr Val Trp Glu Ser Ser 690 695 700 gcg gag gcc tac atc aag gcg agc ttc ccc gag atg cac gca cac atg 2219 Ala Glu Ala Tyr Ile Lys Ala Ser Phe Pro Glu Met His Ala His Met 705 710 715 cgt cgc cac agc gca cct acc act cca cac gga gtg gcc atg ctc acg 2267 Arg Arg His Ser Ala Pro Thr Thr Pro His Gly Val Ala Met Leu Thr 720 725 730 735 agc gac ccg ccc aag ctc aac gcc ttc atc atg gat aaa tca cta ctg 2315 Ser Asp Pro Pro Lys Leu Asn Ala Phe Ile Met Asp Lys Ser Leu Leu 740 745 750 gat tat gag gtg tcc ata gat gcg gac tgc aag ctg ctc acc gtg ggc 2363 Asp Tyr Glu Val Ser Ile Asp Ala Asp Cys Lys Leu Leu Thr Val Gly 755 760 765 aaa ccc ttt gcg atc gag ggc tac ggc ata ggg ctg ccc caa aat tcg 2411 Lys Pro Phe Ala Ile Glu Gly Tyr Gly Ile Gly Leu Pro Gln Asn Ser 770 775 780 ccg ctc acc tcc aac ctg tca gag ttc atc agt agg tac aag tcc tca 2459 Pro Leu Thr Ser Asn Leu Ser Glu Phe Ile Ser Arg Tyr Lys Ser Ser 785 790 795 ggc ttc att gat ctg ctc cat gac aag tgg tac aag atg gtg cct tgc 2507 Gly Phe Ile Asp Leu Leu His Asp Lys Trp Tyr Lys Met Val Pro Cys 800 805 810 815 ggg aag cgg gtg ttc gcc gtg acg gag acg ctg cag atg ggg gtc tac 2555 Gly Lys Arg Val Phe Ala Val Thr Glu Thr Leu Gln Met Gly Val Tyr 820 825 830 cac ttg tca ggg ttg ttt gtc ctg ctg tgc ctc ggg ctg ggc agt gca 2603 His Leu Ser Gly Leu Phe Val Leu Leu Cys Leu Gly Leu Gly Ser Ala 835 840 845 ctt ctc acc tcg ctg ggt gag cac gtc ttc tac cgc ctg gtg ctg ccg 2651 Leu Leu Thr Ser Leu Gly Glu His Val Phe Tyr Arg Leu Val Leu Pro 850 855 860 cgc atc cgc agg ggc aat aag ctg cag tat tgg ctt cac acg agc cag 2699 Arg Ile Arg Arg Gly Asn Lys Leu Gln Tyr Trp Leu His Thr Ser Gln 865 870 875 gaa gat cca ccg agc cct caa cac agg gcc acc aga ggg gca aca gga 2747 Glu Asp Pro Pro Ser Pro Gln His Arg Ala Thr Arg Gly Ala Thr Gly 880 885 890 895 gag ggc aga gca gga gtg cag ggc ccc aag gag gag caa cct gca gcc 2795 Glu Gly Arg Ala Gly Val Gln Gly Pro Lys Glu Glu Gln Pro Ala Ala 900 905 910 gac ggt gcg ggg cgc tgg agg cgg gtg cgc cgg gcc gtg gtg gaa cgg 2843 Asp Gly Ala Gly Arg Trp Arg Arg Val Arg Arg Ala Val Val Glu Arg 915 920 925 gaa cgg cgc gtg cgt ttc ctg ctg gaa cct ggg gag gct ggc ggg gac 2891 Glu Arg Arg Val Arg Phe Leu Leu Glu Pro Gly Glu Ala Gly Gly Asp 930 935 940 cat ccg tgg ctc tgc tcc aat ggg ccc ggg gtg caa gca gaa ctg cgg 2939 His Pro Trp Leu Cys Ser Asn Gly Pro Gly Val Gln Ala Glu Leu Arg 945 950 955 gag ctg gag ctg cgc att gag gct gca cgg gag cgg ctg cgt agt gcg 2987 Glu Leu Glu Leu Arg Ile Glu Ala Ala Arg Glu Arg Leu Arg Ser Ala 960 965 970 975 ctg ctg cgg cga ggg gaa ctg cgg gcc cag ctt ggg gat ggc acc cgg 3035 Leu Leu Arg Arg Gly Glu Leu Arg Ala Gln Leu Gly Asp Gly Thr Arg 980 985 990 ctc agg cca ctg cgc ctg ctg cat gcg gcg ccc gcc gag agc 3077 Leu Arg Pro Leu Arg Leu Leu His Ala Ala Pro Ala Glu Ser 995 1000 1005 tgaggaacta cacggccaca ctgtccacga cagtttattc tatatacaaa cacgactctg 3137 tacactgcaa ttaaatagcg tggaacgtg 3166 <210> SEQ ID NO 8 <211> LENGTH: 1005 <212> TYPE: PRT <213> ORGANISM: Mus musculus <400> SEQUENCE: 8 Met Glu Cys Val Gln Thr Leu Trp Leu Ser Leu Ala Leu Ala Leu Ala 1 5 10 15 Arg Gly Ser Trp Val Val Arg Gly His Pro Gln Pro Cys Gly Val Pro 20 25 30 Thr Arg Ala Gly Ala Ser Val Arg Leu Ala Ala Leu Leu Pro Arg Ala 35 40 45 Pro Ala Ala Arg Ala Arg Val Leu Ala Ala Leu Ala Thr Pro Ser Pro 50 55 60 Arg Leu Pro His Asn Leu Ser Leu Glu Leu Val Ala Val Ala Ser Pro 65 70 75 80 Thr Arg Asp Pro Ala Ser Leu Ala Arg Gly Leu Cys Gln Val Leu Ala 85 90 95 Pro Pro Gly Val Val Ala Ser Ile Thr Phe Pro Glu Ala Arg Pro Glu 100 105 110 Leu Arg Leu Leu Gln Phe Leu Ala Ala Ala Thr Glu Thr Pro Val Leu 115 120 125 Ser Val Leu Arg Arg Glu Val Arg Ala Pro Leu Gly Ala Arg Arg Thr 130 135 140 Pro Phe His Leu Gln Leu Asp Trp Ala Ser Pro Leu Glu Thr Ile Leu 145 150 155 160 Asp Val Leu Val Ser Leu Val Arg Ala His Ala Trp Glu Asp Ile Ala 165 170 175 Leu Val Leu Cys Arg Val Arg Asp Pro Ser Gly Leu Val Thr Leu Trp 180 185 190 Thr Ser Arg Ala Ser Gln Ala Pro Lys Phe Val Leu Asp Leu Ser Gln 195 200 205 Leu Asp Ser Gly Asn Asp Ser Leu Arg Ala Thr Leu Ala Leu Leu Gly 210 215 220 Thr Leu Glu Gly Gly Gly Thr Pro Val Ser Ala Ala Val Leu Leu Gly 225 230 235 240 Cys Ser Thr Ala His Ala His Glu Val Leu Glu Ala Ala Pro Pro Gly 245 250 255 Pro Gln Trp Leu Leu Gly Thr Pro Leu Pro Ala Glu Ala Leu Pro Lys 260 265 270 Thr Gly Leu Pro Pro Gly Val Leu Val Leu Gly Glu Thr Gly Gln Pro 275 280 285 Ser Leu Glu Ala Ala Val His Asp Met Val Glu Leu Val Ala Arg Ala 290 295 300 Leu Ser Ser Met Ala Leu Met His Pro Glu Arg Ala Leu Leu Pro Ala 305 310 315 320 Ala Val Asn Cys Glu Asp Leu Lys Thr Gly Gly Ser Glu Ser Thr Ala 325 330 335 Arg Thr Leu Ala Arg Trp Phe Leu Ser Asn Thr Ser Phe Gln Gly Arg 340 345 350 Thr Gly Ala Val Trp Val Ala Gly Ser Ser Gln Val His Val Ser Arg 355 360 365 His Phe Lys Val Trp Ser Leu Arg Arg Asp Pro Leu Gly Ala Pro Ala 370 375 380 Trp Ala Thr Val Gly Ser Trp Gln Asp Gly Gln Leu Asp Phe Gln Pro 385 390 395 400 Gly Ala Ala Ala Leu Arg Val Pro Ser Pro Ser Gly Thr Gln Ala Arg 405 410 415 Pro Lys Leu Arg Val Val Thr Leu Val Glu His Pro Phe Val Phe Thr 420 425 430 Arg Glu Ser Asp Glu Asp Gly Gln Cys Pro Ala Gly Gln Leu Cys Leu 435 440 445 Asp Pro Gly Thr Asn Asp Ser Ala Arg Leu Asp Ala Leu Phe Thr Ala 450 455 460 Leu Glu Asn Gly Ser Val Pro Arg Thr Leu Arg Arg Cys Cys Tyr Gly 465 470 475 480 Tyr Cys Ile Asp Leu Leu Glu Arg Leu Ala Glu Asp Leu Ala Phe Asp 485 490 495 Phe Glu Leu Tyr Ile Val Gly Asp Gly Lys Tyr Gly Ala Leu Arg Asp 500 505 510 Gly Arg Trp Thr Gly Leu Val Gly Asp Leu Leu Ala Gly Arg Ala His 515 520 525 Met Ala Val Thr Ser Phe Ser Ile Asn Ser Ala Arg Ser Gln Val Val 530 535 540 Asp Phe Thr Ser Pro Phe Phe Ser Thr Ser Leu Gly Ile Met Val Arg 545 550 555 560 Thr Arg Asp Thr Ala Ser Pro Ile Gly Ala Phe Met Trp Pro Leu His 565 570 575 Trp Ser Met Trp Val Gly Val Phe Ala Ala Leu His Leu Thr Ala Leu 580 585 590 Phe Leu Thr Leu Tyr Glu Trp Arg Ser Pro Tyr Gly Leu Thr Pro Arg 595 600 605 Gly Arg Asn Arg Gly Thr Val Phe Ser Tyr Ser Ser Ala Leu Asn Leu 610 615 620 Cys Tyr Ala Ile Leu Phe Gly Arg Thr Val Ser Ser Lys Thr Pro Lys 625 630 635 640 Cys Pro Thr Gly Arg Phe Leu Met Asn Leu Trp Ala Ile Phe Cys Leu 645 650 655 Leu Val Leu Ser Ser Tyr Thr Ala Asn Leu Ala Ala Val Met Val Gly 660 665 670 Asp Lys Thr Phe Glu Glu Leu Ser Gly Ile His Asp Pro Lys Leu His 675 680 685 His Pro Ser Gln Gly Phe Arg Phe Gly Thr Val Trp Glu Ser Ser Ala 690 695 700 Glu Ala Tyr Ile Lys Ala Ser Phe Pro Glu Met His Ala His Met Arg 705 710 715 720 Arg His Ser Ala Pro Thr Thr Pro His Gly Val Ala Met Leu Thr Ser 725 730 735 Asp Pro Pro Lys Leu Asn Ala Phe Ile Met Asp Lys Ser Leu Leu Asp 740 745 750 Tyr Glu Val Ser Ile Asp Ala Asp Cys Lys Leu Leu Thr Val Gly Lys 755 760 765 Pro Phe Ala Ile Glu Gly Tyr Gly Ile Gly Leu Pro Gln Asn Ser Pro 770 775 780 Leu Thr Ser Asn Leu Ser Glu Phe Ile Ser Arg Tyr Lys Ser Ser Gly 785 790 795 800 Phe Ile Asp Leu Leu His Asp Lys Trp Tyr Lys Met Val Pro Cys Gly 805 810 815 Lys Arg Val Phe Ala Val Thr Glu Thr Leu Gln Met Gly Val Tyr His 820 825 830 Leu Ser Gly Leu Phe Val Leu Leu Cys Leu Gly Leu Gly Ser Ala Leu 835 840 845 Leu Thr Ser Leu Gly Glu His Val Phe Tyr Arg Leu Val Leu Pro Arg 850 855 860 Ile Arg Arg Gly Asn Lys Leu Gln Tyr Trp Leu His Thr Ser Gln Glu 865 870 875 880 Asp Pro Pro Ser Pro Gln His Arg Ala Thr Arg Gly Ala Thr Gly Glu 885 890 895 Gly Arg Ala Gly Val Gln Gly Pro Lys Glu Glu Gln Pro Ala Ala Asp 900 905 910 Gly Ala Gly Arg Trp Arg Arg Val Arg Arg Ala Val Val Glu Arg Glu 915 920 925 Arg Arg Val Arg Phe Leu Leu Glu Pro Gly Glu Ala Gly Gly Asp His 930 935 940 Pro Trp Leu Cys Ser Asn Gly Pro Gly Val Gln Ala Glu Leu Arg Glu 945 950 955 960 Leu Glu Leu Arg Ile Glu Ala Ala Arg Glu Arg Leu Arg Ser Ala Leu 965 970 975 Leu Arg Arg Gly Glu Leu Arg Ala Gln Leu Gly Asp Gly Thr Arg Leu 980 985 990 Arg Pro Leu Arg Leu Leu His Ala Ala Pro Ala Glu Ser 995 1000 1005 <210> SEQ ID NO 9 <211> LENGTH: 2691 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <220> FEATURE: <221> NAME/KEY: CDS <222> LOCATION: (1)...(2691) <400> SEQUENCE: 9 atg gag ttt gtg cgg gcg ctg tgg ctg ggc ctg gcg ctg gcg ctg ggg 48 Met Glu Phe Val Arg Ala Leu Trp Leu Gly Leu Ala Leu Ala Leu Gly 1 5 10 15 ccg ggg tcc gcg ggg ggc cac cct cag ccg tgc ggc gtc ctg gcg cgc 96 Pro Gly Ser Ala Gly Gly His Pro Gln Pro Cys Gly Val Leu Ala Arg 20 25 30 ctc ggg ggc tcc gtg cgc ctg ggc gcc ctc ctg ccc cgc gcg cct ctc 144 Leu Gly Gly Ser Val Arg Leu Gly Ala Leu Leu Pro Arg Ala Pro Leu 35 40 45 gcc cgc gcc cgc gcc cgc gcc gcc ctg gcc cgg gcc gcc ctg gcg ccg 192 Ala Arg Ala Arg Ala Arg Ala Ala Leu Ala Arg Ala Ala Leu Ala Pro 50 55 60 cgg ctg ccg cac aac ctg agc ttg gag ctg gtg gtc gcc gcg ccc ccc 240 Arg Leu Pro His Asn Leu Ser Leu Glu Leu Val Val Ala Ala Pro Pro 65 70 75 80 gcc cgc gac ccc gcc tcg ctg acc cgc ggc ctg tgc cag gcg ctg gtg 288 Ala Arg Asp Pro Ala Ser Leu Thr Arg Gly Leu Cys Gln Ala Leu Val 85 90 95 cct ccg ggc gtg gcg gcc ctg ctc gcc ttt ccc gag gct cgg ccc gag 336 Pro Pro Gly Val Ala Ala Leu Leu Ala Phe Pro Glu Ala Arg Pro Glu 100 105 110 ctg ctg cag ctg cac ttc ctg gcg gcg gcc acc gag acc ccc gtg ctc 384 Leu Leu Gln Leu His Phe Leu Ala Ala Ala Thr Glu Thr Pro Val Leu 115 120 125 agc ctg ctg cgg cgg gag gcg cgc gcg ccc ctc gga gcc ccg aac cca 432 Ser Leu Leu Arg Arg Glu Ala Arg Ala Pro Leu Gly Ala Pro Asn Pro 130 135 140 ttc cac ctg cag ctg cac tgg gcc agc ccc ctg gag acg ctg ctg gat 480 Phe His Leu Gln Leu His Trp Ala Ser Pro Leu Glu Thr Leu Leu Asp 145 150 155 160 gtg ctg gtg gcg gtg ctg cag gcg cac gcc tgg gaa gac gtc ggc ctg 528 Val Leu Val Ala Val Leu Gln Ala His Ala Trp Glu Asp Val Gly Leu 165 170 175 gcc ctg tgc cgc act cag gac ccc ggc ggc ctg gtg gcc ctc tgg aca 576 Ala Leu Cys Arg Thr Gln Asp Pro Gly Gly Leu Val Ala Leu Trp Thr 180 185 190 agc cgg gct ggc cgg ccc cca cag ctg gtc ctg gac cta agc cgg cgg 624 Ser Arg Ala Gly Arg Pro Pro Gln Leu Val Leu Asp Leu Ser Arg Arg 195 200 205 gac acg gga gat gca gga ctg cgg gca cgc ctg gcc ccg atg gcg gcg 672 Asp Thr Gly Asp Ala Gly Leu Arg Ala Arg Leu Ala Pro Met Ala Ala 210 215 220 cca gtg ggg ggt gaa gca ccg gta ccc gcg gcg gtc ctc ctc ggc tgt 720 Pro Val Gly Gly Glu Ala Pro Val Pro Ala Ala Val Leu Leu Gly Cys 225 230 235 240 gac atc gcc cgt gcc cgt cgg gtg ctg gag gcc gta cct ccc ggc ccc 768 Asp Ile Ala Arg Ala Arg Arg Val Leu Glu Ala Val Pro Pro Gly Pro 245 250 255 cac tgg ctg ttg ggg aca cca ctg ccg ccc aag gcc ctg ccc acc gcg 816 His Trp Leu Leu Gly Thr Pro Leu Pro Pro Lys Ala Leu Pro Thr Ala 260 265 270 ggg ctg cca cca ggg ctg ctg gcg ctg ggc gag gtg gca cga ccc ccg 864 Gly Leu Pro Pro Gly Leu Leu Ala Leu Gly Glu Val Ala Arg Pro Pro 275 280 285 ctg gag gcc gcc atc cat gac att gtg caa ctg gtg gcc cgg gcg ctg 912 Leu Glu Ala Ala Ile His Asp Ile Val Gln Leu Val Ala Arg Ala Leu 290 295 300 ggc agt gcg gcc cag gtg cag ccg aag cga gcc ctc ctc ccc gcc ccg 960 Gly Ser Ala Ala Gln Val Gln Pro Lys Arg Ala Leu Leu Pro Ala Pro 305 310 315 320 gtc aac tgc ggg gac ctg cag ccg gcc ggg ccc gag tcc ccg ggg cgc 1008 Val Asn Cys Gly Asp Leu Gln Pro Ala Gly Pro Glu Ser Pro Gly Arg 325 330 335 ttc ttg gcc aac acg tcc ttc cag ggc cgc acg ggc ccc gtg tgg gtg 1056 Phe Leu Ala Asn Thr Ser Phe Gln Gly Arg Thr Gly Pro Val Trp Val 340 345 350 aca ggc agc tcc cag gta cac atg tct cgg cac ttt aag gtg tgg agc 1104 Thr Gly Ser Ser Gln Val His Met Ser Arg His Phe Lys Val Trp Ser 355 360 365 ctt cgc cgg gac cca cgg ggc gcc ccg gcc tgg gcc acg gtg ggc agc 1152 Leu Arg Arg Asp Pro Arg Gly Ala Pro Ala Trp Ala Thr Val Gly Ser 370 375 380 tgg cgg gac ggc cag ctg gac ttg gaa ccg gga ggt gcc tct gca cgg 1200 Trp Arg Asp Gly Gln Leu Asp Leu Glu Pro Gly Gly Ala Ser Ala Arg 385 390 395 400 ccc ccg ccc cca cag ggt gcc cag gtc tgg ccc aag ctg cgt gtg gta 1248 Pro Pro Pro Pro Gln Gly Ala Gln Val Trp Pro Lys Leu Arg Val Val 405 410 415 acg ctg ttg gaa cac cca ttt gtg ttt gcc cgt gat cca gac gaa gac 1296 Thr Leu Leu Glu His Pro Phe Val Phe Ala Arg Asp Pro Asp Glu Asp 420 425 430 ggg cag tgc cca gcg ggg cag ctg tgc ctg gac cct ggc acc aac gac 1344 Gly Gln Cys Pro Ala Gly Gln Leu Cys Leu Asp Pro Gly Thr Asn Asp 435 440 445 tcg gcc acc ctg gac gca ctg ttc gcc gcg ctg gcc aac ggc tca gcg 1392 Ser Ala Thr Leu Asp Ala Leu Phe Ala Ala Leu Ala Asn Gly Ser Ala 450 455 460 ccc cgt gcc ctg cgc aag tgc tgc tac ggc tac tgc att gac ctg ctg 1440 Pro Arg Ala Leu Arg Lys Cys Cys Tyr Gly Tyr Cys Ile Asp Leu Leu 465 470 475 480 gag cgg ctg gcg gag gac acg ccc ttc gac ttc gag ctg tac ctc gtg 1488 Glu Arg Leu Ala Glu Asp Thr Pro Phe Asp Phe Glu Leu Tyr Leu Val 485 490 495 ggt gac ggc aag tac ggc gcc ctg cgg gac ggc cgc tgg acc ggc ctg 1536 Gly Asp Gly Lys Tyr Gly Ala Leu Arg Asp Gly Arg Trp Thr Gly Leu 500 505 510 gtc ggg gac ctg ctg gcc ggc cgg gcc cac atg gcg gtc acc agc ttc 1584 Val Gly Asp Leu Leu Ala Gly Arg Ala His Met Ala Val Thr Ser Phe 515 520 525 agt atc aac tcc gcc cgc tca cag gtg gtg gac ttc acc agc ccc ttc 1632 Ser Ile Asn Ser Ala Arg Ser Gln Val Val Asp Phe Thr Ser Pro Phe 530 535 540 ttc tcc acc agc ctg ggc atc atg gtg cgg gca cgg gac acg gcc tca 1680 Phe Ser Thr Ser Leu Gly Ile Met Val Arg Ala Arg Asp Thr Ala Ser 545 550 555 560 ccc atc ggt gcc ttt atg tgg ccc ctg cac tgg tcc acg tgg ctg ggc 1728 Pro Ile Gly Ala Phe Met Trp Pro Leu His Trp Ser Thr Trp Leu Gly 565 570 575 gtc ttt gcg gcc ctg cac ctc acc gcg ctc ttc ctc acc gtg tac gag 1776 Val Phe Ala Ala Leu His Leu Thr Ala Leu Phe Leu Thr Val Tyr Glu 580 585 590 tgg cgt agc ccc tac ggc ctc acg cca cgt ggc cgc aac cgc agc acc 1824 Trp Arg Ser Pro Tyr Gly Leu Thr Pro Arg Gly Arg Asn Arg Ser Thr 595 600 605 gtc ttc tcc tac tcc tca gcc ctc aac ctg tgc tac gcc atc ctc ttc 1872 Val Phe Ser Tyr Ser Ser Ala Leu Asn Leu Cys Tyr Ala Ile Leu Phe 610 615 620 aga cgc acc gtg tcc agc aag acg ccc aag tgc ccc acg ggc cgc ctg 1920 Arg Arg Thr Val Ser Ser Lys Thr Pro Lys Cys Pro Thr Gly Arg Leu 625 630 635 640 ctc atg aac ctc tgg gcc atc ttc tgc ctg ctg gtg ctg tcc agc tac 1968 Leu Met Asn Leu Trp Ala Ile Phe Cys Leu Leu Val Leu Ser Ser Tyr 645 650 655 acg gcc aac ctg gct gcc gtc atg gtc ggg gac aag acc ttc gag gag 2016 Thr Ala Asn Leu Ala Ala Val Met Val Gly Asp Lys Thr Phe Glu Glu 660 665 670 ctg tcg ggg atc cac gac ccc aag ctg cac cac ccg gcg cag ggc ttc 2064 Leu Ser Gly Ile His Asp Pro Lys Leu His His Pro Ala Gln Gly Phe 675 680 685 cgc ttc ggc acc gtg tgg gag agc agc gcc gag gcg tac atc aag aag 2112 Arg Phe Gly Thr Val Trp Glu Ser Ser Ala Glu Ala Tyr Ile Lys Lys 690 695 700 agc ttc ccc gac atg cac gca cac atg cgg cgc cac agc gcg ccc acc 2160 Ser Phe Pro Asp Met His Ala His Met Arg Arg His Ser Ala Pro Thr 705 710 715 720 acg ccc cgc ggc gtc gcc atg ctc acg agc gac ccc ccc aag ctc aac 2208 Thr Pro Arg Gly Val Ala Met Leu Thr Ser Asp Pro Pro Lys Leu Asn 725 730 735 gcc ttc atc atg gac aag tcg ctc ctg gac tac gag gtc tcc atc gac 2256 Ala Phe Ile Met Asp Lys Ser Leu Leu Asp Tyr Glu Val Ser Ile Asp 740 745 750 gcc gac tgc aaa ctg ctg acc gtg gga aag ccc ttc gcc att gag ggc 2304 Ala Asp Cys Lys Leu Leu Thr Val Gly Lys Pro Phe Ala Ile Glu Gly 755 760 765 tat ggg atc gga ctg ccc cag aac tcg ccg ctc acc tcc aac ctg tcc 2352 Tyr Gly Ile Gly Leu Pro Gln Asn Ser Pro Leu Thr Ser Asn Leu Ser 770 775 780 gag ttc atc agc cgc tac aag tcc tcc ggc ttc atc gac ctg ctc cac 2400 Glu Phe Ile Ser Arg Tyr Lys Ser Ser Gly Phe Ile Asp Leu Leu His 785 790 795 800 gac aag tgg tac aag atg gtg cct tgc ggc aag cgg gtc ttt gcg gtt 2448 Asp Lys Trp Tyr Lys Met Val Pro Cys Gly Lys Arg Val Phe Ala Val 805 810 815 aca gag acc ctg cag atg agc atc tac cac ttc gcg ggc ctc ttc gtg 2496 Thr Glu Thr Leu Gln Met Ser Ile Tyr His Phe Ala Gly Leu Phe Val 820 825 830 ttg ctg tgc ctg ggc ctg ggc agc gct ctg ctc agc tcg ctg ggc gag 2544 Leu Leu Cys Leu Gly Leu Gly Ser Ala Leu Leu Ser Ser Leu Gly Glu 835 840 845 cac gcc ttc ttc cgc ctg gcg ctg ccg cgc atc cgc aag ggg agc agg 2592 His Ala Phe Phe Arg Leu Ala Leu Pro Arg Ile Arg Lys Gly Ser Arg 850 855 860 ctg cag tac tgg ctg cac acc agc cag aaa atc cac cgc gcc ctc aac 2640 Leu Gln Tyr Trp Leu His Thr Ser Gln Lys Ile His Arg Ala Leu Asn 865 870 875 880 acg gag cca cca gag ggg tcg aag gag gag acg gca gag gcg gag ccc 2688 Thr Glu Pro Pro Glu Gly Ser Lys Glu Glu Thr Ala Glu Ala Glu Pro 885 890 895 agg 2691 Arg <210> SEQ ID NO 10 <211> LENGTH: 897 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE: 10 Met Glu Phe Val Arg Ala Leu Trp Leu Gly Leu Ala Leu Ala Leu Gly 1 5 10 15 Pro Gly Ser Ala Gly Gly His Pro Gln Pro Cys Gly Val Leu Ala Arg 20 25 30 Leu Gly Gly Ser Val Arg Leu Gly Ala Leu Leu Pro Arg Ala Pro Leu 35 40 45 Ala Arg Ala Arg Ala Arg Ala Ala Leu Ala Arg Ala Ala Leu Ala Pro 50 55 60 Arg Leu Pro His Asn Leu Ser Leu Glu Leu Val Val Ala Ala Pro Pro 65 70 75 80 Ala Arg Asp Pro Ala Ser Leu Thr Arg Gly Leu Cys Gln Ala Leu Val 85 90 95 Pro Pro Gly Val Ala Ala Leu Leu Ala Phe Pro Glu Ala Arg Pro Glu 100 105 110 Leu Leu Gln Leu His Phe Leu Ala Ala Ala Thr Glu Thr Pro Val Leu 115 120 125 Ser Leu Leu Arg Arg Glu Ala Arg Ala Pro Leu Gly Ala Pro Asn Pro 130 135 140 Phe His Leu Gln Leu His Trp Ala Ser Pro Leu Glu Thr Leu Leu Asp 145 150 155 160 Val Leu Val Ala Val Leu Gln Ala His Ala Trp Glu Asp Val Gly Leu 165 170 175 Ala Leu Cys Arg Thr Gln Asp Pro Gly Gly Leu Val Ala Leu Trp Thr 180 185 190 Ser Arg Ala Gly Arg Pro Pro Gln Leu Val Leu Asp Leu Ser Arg Arg 195 200 205 Asp Thr Gly Asp Ala Gly Leu Arg Ala Arg Leu Ala Pro Met Ala Ala 210 215 220 Pro Val Gly Gly Glu Ala Pro Val Pro Ala Ala Val Leu Leu Gly Cys 225 230 235 240 Asp Ile Ala Arg Ala Arg Arg Val Leu Glu Ala Val Pro Pro Gly Pro 245 250 255 His Trp Leu Leu Gly Thr Pro Leu Pro Pro Lys Ala Leu Pro Thr Ala 260 265 270 Gly Leu Pro Pro Gly Leu Leu Ala Leu Gly Glu Val Ala Arg Pro Pro 275 280 285 Leu Glu Ala Ala Ile His Asp Ile Val Gln Leu Val Ala Arg Ala Leu 290 295 300 Gly Ser Ala Ala Gln Val Gln Pro Lys Arg Ala Leu Leu Pro Ala Pro 305 310 315 320 Val Asn Cys Gly Asp Leu Gln Pro Ala Gly Pro Glu Ser Pro Gly Arg 325 330 335 Phe Leu Ala Asn Thr Ser Phe Gln Gly Arg Thr Gly Pro Val Trp Val 340 345 350 Thr Gly Ser Ser Gln Val His Met Ser Arg His Phe Lys Val Trp Ser 355 360 365 Leu Arg Arg Asp Pro Arg Gly Ala Pro Ala Trp Ala Thr Val Gly Ser 370 375 380 Trp Arg Asp Gly Gln Leu Asp Leu Glu Pro Gly Gly Ala Ser Ala Arg 385 390 395 400 Pro Pro Pro Pro Gln Gly Ala Gln Val Trp Pro Lys Leu Arg Val Val 405 410 415 Thr Leu Leu Glu His Pro Phe Val Phe Ala Arg Asp Pro Asp Glu Asp 420 425 430 Gly Gln Cys Pro Ala Gly Gln Leu Cys Leu Asp Pro Gly Thr Asn Asp 435 440 445 Ser Ala Thr Leu Asp Ala Leu Phe Ala Ala Leu Ala Asn Gly Ser Ala 450 455 460 Pro Arg Ala Leu Arg Lys Cys Cys Tyr Gly Tyr Cys Ile Asp Leu Leu 465 470 475 480 Glu Arg Leu Ala Glu Asp Thr Pro Phe Asp Phe Glu Leu Tyr Leu Val 485 490 495 Gly Asp Gly Lys Tyr Gly Ala Leu Arg Asp Gly Arg Trp Thr Gly Leu 500 505 510 Val Gly Asp Leu Leu Ala Gly Arg Ala His Met Ala Val Thr Ser Phe 515 520 525 Ser Ile Asn Ser Ala Arg Ser Gln Val Val Asp Phe Thr Ser Pro Phe 530 535 540 Phe Ser Thr Ser Leu Gly Ile Met Val Arg Ala Arg Asp Thr Ala Ser 545 550 555 560 Pro Ile Gly Ala Phe Met Trp Pro Leu His Trp Ser Thr Trp Leu Gly 565 570 575 Val Phe Ala Ala Leu His Leu Thr Ala Leu Phe Leu Thr Val Tyr Glu 580 585 590 Trp Arg Ser Pro Tyr Gly Leu Thr Pro Arg Gly Arg Asn Arg Ser Thr 595 600 605 Val Phe Ser Tyr Ser Ser Ala Leu Asn Leu Cys Tyr Ala Ile Leu Phe 610 615 620 Arg Arg Thr Val Ser Ser Lys Thr Pro Lys Cys Pro Thr Gly Arg Leu 625 630 635 640 Leu Met Asn Leu Trp Ala Ile Phe Cys Leu Leu Val Leu Ser Ser Tyr 645 650 655 Thr Ala Asn Leu Ala Ala Val Met Val Gly Asp Lys Thr Phe Glu Glu 660 665 670 Leu Ser Gly Ile His Asp Pro Lys Leu His His Pro Ala Gln Gly Phe 675 680 685 Arg Phe Gly Thr Val Trp Glu Ser Ser Ala Glu Ala Tyr Ile Lys Lys 690 695 700 Ser Phe Pro Asp Met His Ala His Met Arg Arg His Ser Ala Pro Thr 705 710 715 720 Thr Pro Arg Gly Val Ala Met Leu Thr Ser Asp Pro Pro Lys Leu Asn 725 730 735 Ala Phe Ile Met Asp Lys Ser Leu Leu Asp Tyr Glu Val Ser Ile Asp 740 745 750 Ala Asp Cys Lys Leu Leu Thr Val Gly Lys Pro Phe Ala Ile Glu Gly 755 760 765 Tyr Gly Ile Gly Leu Pro Gln Asn Ser Pro Leu Thr Ser Asn Leu Ser 770 775 780 Glu Phe Ile Ser Arg Tyr Lys Ser Ser Gly Phe Ile Asp Leu Leu His 785 790 795 800 Asp Lys Trp Tyr Lys Met Val Pro Cys Gly Lys Arg Val Phe Ala Val 805 810 815 Thr Glu Thr Leu Gln Met Ser Ile Tyr His Phe Ala Gly Leu Phe Val 820 825 830 Leu Leu Cys Leu Gly Leu Gly Ser Ala Leu Leu Ser Ser Leu Gly Glu 835 840 845 His Ala Phe Phe Arg Leu Ala Leu Pro Arg Ile Arg Lys Gly Ser Arg 850 855 860 Leu Gln Tyr Trp Leu His Thr Ser Gln Lys Ile His Arg Ala Leu Asn 865 870 875 880 Thr Glu Pro Pro Glu Gly Ser Lys Glu Glu Thr Ala Glu Ala Glu Pro 885 890 895 Arg <210> SEQ ID NO 11 <211> LENGTH: 1786 <212> TYPE: DNA <213> ORGANISM: Mus musculus <220> FEATURE: <221> NAME/KEY: CDS <222> LOCATION: (2)...(1444) <400> SEQUENCE: 11 c atc aac tgc tta gag cat gtt cga gat cga tgg ccg cgg gag ggt gtc 49 Ile Asn Cys Leu Glu His Val Arg Asp Arg Trp Pro Arg Glu Gly Val 1 5 10 15 ctg cgg gtg gag gtg cgc cac aac tcg agc cgg gca ccg gtg atc ctg 97 Leu Arg Val Glu Val Arg His Asn Ser Ser Arg Ala Pro Val Ile Leu 20 25 30 cag ttc tgt gat ggg ggc ctc ggt ggc ctg gag ctg gaa ccc ggg ggc 145 Gln Phe Cys Asp Gly Gly Leu Gly Gly Leu Glu Leu Glu Pro Gly Gly 35 40 45 ctg gag ctg gag gag gag gag ctc aca gtg gag atg ttc acc aac agc 193 Leu Glu Leu Glu Glu Glu Glu Leu Thr Val Glu Met Phe Thr Asn Ser 50 55 60 tcc atc aag ttt gag ctg gac att gag ccc aag gtg ttc aag cca cag 241 Ser Ile Lys Phe Glu Leu Asp Ile Glu Pro Lys Val Phe Lys Pro Gln 65 70 75 80 agc ggt gca gat gcc ctg aac gac agc cag gac ttc cct ttt cct gag 289 Ser Gly Ala Asp Ala Leu Asn Asp Ser Gln Asp Phe Pro Phe Pro Glu 85 90 95 acg cca gca aaa gtg tgg cca cag gat gag tac att gtg gag tac tca 337 Thr Pro Ala Lys Val Trp Pro Gln Asp Glu Tyr Ile Val Glu Tyr Ser 100 105 110 ctg gaa tat ggc ttc ctg cgg cta tcc caa gcc aca cgc cag cgt ctg 385 Leu Glu Tyr Gly Phe Leu Arg Leu Ser Gln Ala Thr Arg Gln Arg Leu 115 120 125 agc att cct gtc atg gtg gtc acc cta gac ccc acg cgg gac cag tgc 433 Ser Ile Pro Val Met Val Val Thr Leu Asp Pro Thr Arg Asp Gln Cys 130 135 140 ttt ggg gac cgc ttc agc cgc cta ttg ctg gat gag ttc ctg ggc tat 481 Phe Gly Asp Arg Phe Ser Arg Leu Leu Leu Asp Glu Phe Leu Gly Tyr 145 150 155 160 gat gac atc ctc atg tcc agt gta aag ggt ctg gca gag aac gag gag 529 Asp Asp Ile Leu Met Ser Ser Val Lys Gly Leu Ala Glu Asn Glu Glu 165 170 175 aac aaa ggc ttc ttg agg aat gtg gtc tct ggg gag cac tac cgc ttt 577 Asn Lys Gly Phe Leu Arg Asn Val Val Ser Gly Glu His Tyr Arg Phe 180 185 190 gtc agc atg tgg atg gcg cgc aca tcc tac ctg gcg gcc ttt gtc atc 625 Val Ser Met Trp Met Ala Arg Thr Ser Tyr Leu Ala Ala Phe Val Ile 195 200 205 atg gtc atc ttt acc ctc agc gtg tcc atg ctg ttg cga tac tcg cac 673 Met Val Ile Phe Thr Leu Ser Val Ser Met Leu Leu Arg Tyr Ser His 210 215 220 cac cag atc ttc gtc ttc atc gtg gac ctg ctg cag atg ctg gag atg 721 His Gln Ile Phe Val Phe Ile Val Asp Leu Leu Gln Met Leu Glu Met 225 230 235 240 aac atg gcc atc gcc ttc ccc gca gcg ccc ttg ctg acc gtc atc ctg 769 Asn Met Ala Ile Ala Phe Pro Ala Ala Pro Leu Leu Thr Val Ile Leu 245 250 255 gct ctc gtc ggg atg gaa gcc atc atg tct gag ttc ttc aat gat acc 817 Ala Leu Val Gly Met Glu Ala Ile Met Ser Glu Phe Phe Asn Asp Thr 260 265 270 acc acg gcc ttc tac atc atc ctc act gtg tgg ctg gcc gac cag tat 865 Thr Thr Ala Phe Tyr Ile Ile Leu Thr Val Trp Leu Ala Asp Gln Tyr 275 280 285 gat gcc atc tgc tgc cac acc aac acc agc aag cgg cac tgg ctg agg 913 Asp Ala Ile Cys Cys His Thr Asn Thr Ser Lys Arg His Trp Leu Arg 290 295 300 ttc ttc tac ctc tac cac ttc gcc ttc tat gcc tac cac tac cgc ttt 961 Phe Phe Tyr Leu Tyr His Phe Ala Phe Tyr Ala Tyr His Tyr Arg Phe 305 310 315 320 aac ggg cag tac agc agc ctg gcc ctg gtc acc tcc tgg ctc ttc atc 1009 Asn Gly Gln Tyr Ser Ser Leu Ala Leu Val Thr Ser Trp Leu Phe Ile 325 330 335 cag cat tcc atg atc tac ttc ttc cac cac tat gag ttg ccc gcc atc 1057 Gln His Ser Met Ile Tyr Phe Phe His His Tyr Glu Leu Pro Ala Ile 340 345 350 ctg cag cag atc cga atc cag gag atg ctg ctt cag acg cca ccg ctg 1105 Leu Gln Gln Ile Arg Ile Gln Glu Met Leu Leu Gln Thr Pro Pro Leu 355 360 365 ggc ccc ggg acc ccc acg gcg ctg cct gac gac ctc aac aac aac tct 1153 Gly Pro Gly Thr Pro Thr Ala Leu Pro Asp Asp Leu Asn Asn Asn Ser 370 375 380 ggc tcc cct gcc act ccg gat ccc agc cct ccc ctc gcg ctg ggc ccc 1201 Gly Ser Pro Ala Thr Pro Asp Pro Ser Pro Pro Leu Ala Leu Gly Pro 385 390 395 400 agc tcc agc ccc gcg ccc act ggc ggg gca tct ggg cct ggc tca ctg 1249 Ser Ser Ser Pro Ala Pro Thr Gly Gly Ala Ser Gly Pro Gly Ser Leu 405 410 415 ggc gct ggg gcc tca gta tcc ggc agt gac cta ggt tgg gtg gcc gag 1297 Gly Ala Gly Ala Ser Val Ser Gly Ser Asp Leu Gly Trp Val Ala Glu 420 425 430 acc gcc gcc atc atc tct gac gca tcc ttc ctg tcg ggg ctg agc gcc 1345 Thr Ala Ala Ile Ile Ser Asp Ala Ser Phe Leu Ser Gly Leu Ser Ala 435 440 445 tct ctc ctg gag cgg cgg cca aca gcc cct agt acc ccg gac agc tca 1393 Ser Leu Leu Glu Arg Arg Pro Thr Ala Pro Ser Thr Pro Asp Ser Ser 450 455 460 cga cct gac cct ggg gtc cct ctg gag gac gca ccc gcc cct gcc ggg 1441 Arg Pro Asp Pro Gly Val Pro Leu Glu Asp Ala Pro Ala Pro Ala Gly 465 470 475 480 tcc tgagaccggt gtcgtgcccg gctcccagtg gagccgcggc ccgagcccaa 1494 Ser gagagctgtg gtctgcaggg agaggggctg gtggcgaagg ttctggaagc ggccgggaca 1554 gggcggcgat gggcacgagg ccatcggccg cctggtgctc cccagtgcct tccacacggc 1614 gcccgcacgg ggccgagcgc ccgggcccgg actagcaggt ggtggctcac gatgcggggc 1674 acacgttcca cgctatttaa ttgcagtgta cagagtcgtg tttgtatata gaataaactg 1734 tcgtggacag tgaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aa 1786 <210> SEQ ID NO 12 <211> LENGTH: 481 <212> TYPE: PRT <213> ORGANISM: Mus musculus <400> SEQUENCE: 12 Ile Asn Cys Leu Glu His Val Arg Asp Arg Trp Pro Arg Glu Gly Val 1 5 10 15 Leu Arg Val Glu Val Arg His Asn Ser Ser Arg Ala Pro Val Ile Leu 20 25 30 Gln Phe Cys Asp Gly Gly Leu Gly Gly Leu Glu Leu Glu Pro Gly Gly 35 40 45 Leu Glu Leu Glu Glu Glu Glu Leu Thr Val Glu Met Phe Thr Asn Ser 50 55 60 Ser Ile Lys Phe Glu Leu Asp Ile Glu Pro Lys Val Phe Lys Pro Gln 65 70 75 80 Ser Gly Ala Asp Ala Leu Asn Asp Ser Gln Asp Phe Pro Phe Pro Glu 85 90 95 Thr Pro Ala Lys Val Trp Pro Gln Asp Glu Tyr Ile Val Glu Tyr Ser 100 105 110 Leu Glu Tyr Gly Phe Leu Arg Leu Ser Gln Ala Thr Arg Gln Arg Leu 115 120 125 Ser Ile Pro Val Met Val Val Thr Leu Asp Pro Thr Arg Asp Gln Cys 130 135 140 Phe Gly Asp Arg Phe Ser Arg Leu Leu Leu Asp Glu Phe Leu Gly Tyr 145 150 155 160 Asp Asp Ile Leu Met Ser Ser Val Lys Gly Leu Ala Glu Asn Glu Glu 165 170 175 Asn Lys Gly Phe Leu Arg Asn Val Val Ser Gly Glu His Tyr Arg Phe 180 185 190 Val Ser Met Trp Met Ala Arg Thr Ser Tyr Leu Ala Ala Phe Val Ile 195 200 205 Met Val Ile Phe Thr Leu Ser Val Ser Met Leu Leu Arg Tyr Ser His 210 215 220 His Gln Ile Phe Val Phe Ile Val Asp Leu Leu Gln Met Leu Glu Met 225 230 235 240 Asn Met Ala Ile Ala Phe Pro Ala Ala Pro Leu Leu Thr Val Ile Leu 245 250 255 Ala Leu Val Gly Met Glu Ala Ile Met Ser Glu Phe Phe Asn Asp Thr 260 265 270 Thr Thr Ala Phe Tyr Ile Ile Leu Thr Val Trp Leu Ala Asp Gln Tyr 275 280 285 Asp Ala Ile Cys Cys His Thr Asn Thr Ser Lys Arg His Trp Leu Arg 290 295 300 Phe Phe Tyr Leu Tyr His Phe Ala Phe Tyr Ala Tyr His Tyr Arg Phe 305 310 315 320 Asn Gly Gln Tyr Ser Ser Leu Ala Leu Val Thr Ser Trp Leu Phe Ile 325 330 335 Gln His Ser Met Ile Tyr Phe Phe His His Tyr Glu Leu Pro Ala Ile 340 345 350 Leu Gln Gln Ile Arg Ile Gln Glu Met Leu Leu Gln Thr Pro Pro Leu 355 360 365 Gly Pro Gly Thr Pro Thr Ala Leu Pro Asp Asp Leu Asn Asn Asn Ser 370 375 380 Gly Ser Pro Ala Thr Pro Asp Pro Ser Pro Pro Leu Ala Leu Gly Pro 385 390 395 400 Ser Ser Ser Pro Ala Pro Thr Gly Gly Ala Ser Gly Pro Gly Ser Leu 405 410 415 Gly Ala Gly Ala Ser Val Ser Gly Ser Asp Leu Gly Trp Val Ala Glu 420 425 430 Thr Ala Ala Ile Ile Ser Asp Ala Ser Phe Leu Ser Gly Leu Ser Ala 435 440 445 Ser Leu Leu Glu Arg Arg Pro Thr Ala Pro Ser Thr Pro Asp Ser Ser 450 455 460 Arg Pro Asp Pro Gly Val Pro Leu Glu Asp Ala Pro Ala Pro Ala Gly 465 470 475 480 Ser <210> SEQ ID NO 13 <211> LENGTH: 579 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 13 ccacgcgtcc gatcttgtac cacttgtcgt ggagcaggtc gatgaagccg gaggacttgt 60 agcggctgat gaactcggac aggttggagg aggtccccag agaccccggc gccccgcctc 120 gcccaggtgc ctctcaccct caatggcgaa gggctttccc acggtcagca gtttgcagtc 180 ggcgtcgatg gagacctcgt agtccaggag cgacttgtcc atgatgaagg cgttgagctt 240 gggggggtcg ctcctgctgg ggccgggggc gggggtcagc catcgccccg cccaccccac 300 gccccgcccc cgcctcaccc cgcgcccggg ctcacgtgag catggcgacy ccgcggggcg 360 tggtgggcgc gctgtggcgc cgcatgtgtg cgtgcatgtc ggggaagctc ttcttgatgt 420 acgcctcggc gctgttctcc cacacggtgc cgaagcggaa gccctgcgcc gggtggtgca 480 gctgcgcggg ggaccccgtc agcgcctctg ctgcccctca ggacccctga ccattgaggg 540 gcgcgccgtt ctccggggtg gggccgtcct gggcacttg 579 <210> SEQ ID NO 14 <211> LENGTH: 938 <212> TYPE: PRT <213> ORGANISM: Rattus sp. <400> SEQUENCE: 14 Met Ser Thr Met His Leu Leu Thr Phe Ala Leu Leu Phe Ser Cys Ser 1 5 10 15 Phe Ala Arg Ala Ala Cys Asp Pro Lys Ile Val Asn Ile Gly Ala Val 20 25 30 Leu Ser Thr Arg Lys His Glu Gln Met Phe Arg Glu Ala Val Asn Gln 35 40 45 Ala Asn Lys Arg His Gly Ser Trp Lys Ile Gln Leu Asn Ala Thr Ser 50 55 60 Val Thr His Lys Pro Asn Ala Ile Gln Met Ala Leu Ser Val Cys Glu 65 70 75 80 Asp Leu Ile Ser Ser Gln Val Tyr Ala Ile Leu Val Ser His Pro Pro 85 90 95 Thr Pro Asn Asp His Phe Thr Pro Thr Pro Val Ser Tyr Thr Ala Gly 100 105 110 Phe Tyr Arg Ile Pro Val Leu Gly Leu Thr Thr Arg Met Ser Ile Tyr 115 120 125 Ser Asp Lys Ser Ile His Leu Ser Phe Leu Arg Thr Val Pro Pro Tyr 130 135 140 Ser His Gln Ser Ser Val Trp Phe Glu Met Met Arg Val Tyr Asn Trp 145 150 155 160 Asn His Ile Ile Leu Leu Val Ser Asp Asp His Glu Gly Arg Ala Ala 165 170 175 Gln Lys Arg Leu Glu Thr Leu Leu Glu Glu Arg Glu Ser Lys Ala Glu 180 185 190 Lys Val Leu Gln Phe Asp Pro Gly Thr Lys Asn Val Thr Ala Leu Leu 195 200 205 Met Glu Ala Arg Glu Leu Glu Ala Arg Val Ile Ile Leu Ser Ala Ser 210 215 220 Glu Asp Asp Ala Ala Thr Val Tyr Arg Ala Ala Ala Met Leu Asn Met 225 230 235 240 Thr Gly Ser Gly Tyr Val Trp Leu Val Gly Glu Arg Glu Ile Ser Gly 245 250 255 Asn Ala Leu Arg Tyr Ala Pro Asp Gly Ile Ile Gly Leu Gln Leu Ile 260 265 270 Asn Gly Lys Asn Glu Ser Ala His Ile Ser Asp Ala Val Gly Val Val 275 280 285 Ala Gln Ala Val His Glu Leu Leu Glu Lys Glu Asn Ile Thr Asp Pro 290 295 300 Pro Arg Gly Cys Val Gly Asn Thr Asn Ile Trp Lys Thr Gly Pro Leu 305 310 315 320 Phe Lys Arg Val Leu Met Ser Ser Lys Tyr Ala Asp Gly Val Thr Gly 325 330 335 Arg Val Glu Phe Asn Glu Asp Gly Asp Arg Lys Phe Ala Asn Tyr Ser 340 345 350 Ile Met Asn Leu Gln Asn Arg Lys Leu Val Gln Val Gly Ile Tyr Asn 355 360 365 Gly Thr His Val Ile Pro Asn Asp Arg Lys Ile Ile Trp Pro Gly Gly 370 375 380 Glu Thr Glu Lys Pro Arg Gly Tyr Gln Met Ser Thr Arg Leu Lys Ile 385 390 395 400 Val Thr Ile His Gln Glu Pro Phe Val Tyr Val Lys Pro Thr Met Ser 405 410 415 Asp Gly Thr Cys Lys Glu Glu Phe Thr Val Asn Gly Asp Pro Val Lys 420 425 430 Lys Val Ile Cys Thr Gly Pro Asn Asp Thr Ser Pro Gly Ser Pro Arg 435 440 445 His Thr Val Pro Gln Cys Cys Tyr Gly Phe Cys Ile Asp Leu Leu Ile 450 455 460 Lys Leu Ala Arg Thr Met Asn Phe Thr Tyr Glu Val His Leu Val Ala 465 470 475 480 Asp Gly Lys Phe Gly Thr Gln Glu Arg Val Asn Asn Ser Asn Lys Lys 485 490 495 Glu Trp Asn Gly Met Met Gly Glu Leu Leu Ser Gly Gln Ala Asp Met 500 505 510 Ile Val Ala Pro Leu Thr Ile Asn Asn Glu Arg Ala Gln Tyr Ile Glu 515 520 525 Phe Ser Lys Pro Phe Lys Tyr Gln Gly Leu Thr Ile Leu Val Lys Lys 530 535 540 Glu Ile Pro Arg Ser Thr Leu Asp Ser Phe Met Gln Pro Phe Gln Ser 545 550 555 560 Thr Leu Trp Leu Leu Val Gly Leu Ser Val His Val Val Ala Val Met 565 570 575 Leu Tyr Leu Leu Asp Arg Phe Ser Pro Phe Gly Arg Phe Lys Val Asn 580 585 590 Ser Glu Glu Glu Glu Glu Asp Ala Leu Thr Leu Ser Ser Ala Met Trp 595 600 605 Phe Ser Trp Gly Val Leu Leu Asn Ser Gly Ile Gly Glu Gly Ala Pro 610 615 620 Arg Ser Phe Ser Ala Arg Ile Leu Gly Met Val Trp Ala Gly Phe Ala 625 630 635 640 Met Ile Ile Val Ala Ser Tyr Thr Ala Asn Leu Ala Ala Phe Leu Val 645 650 655 Leu Asp Arg Pro Glu Glu Arg Ile Thr Gly Ile Asn Asp Pro Arg Leu 660 665 670 Arg Asn Pro Ser Asp Lys Phe Ile Tyr Ala Thr Val Lys Gln Ser Ser 675 680 685 Val Asp Ile Tyr Phe Arg Arg Gln Val Glu Leu Ser Thr Met Tyr Arg 690 695 700 His Met Glu Lys His Asn Tyr Glu Ser Ala Ala Glu Ala Ile Gln Ala 705 710 715 720 Val Arg Asp Asn Lys Leu His Ala Phe Ile Trp Asp Ser Ala Val Leu 725 730 735 Glu Phe Glu Ala Ser Gln Lys Cys Asp Leu Val Thr Thr Gly Glu Leu 740 745 750 Phe Phe Arg Ser Gly Phe Gly Ile Gly Met Arg Lys Asp Ser Pro Trp 755 760 765 Lys Gln Asn Val Ser Leu Ser Ile Leu Lys Ser His Glu Asn Gly Phe 770 775 780 Met Glu Asp Leu Asp Lys Thr Trp Val Arg Tyr Gln Glu Cys Asp Ser 785 790 795 800 Arg Ser Asn Ala Pro Ala Thr Leu Thr Phe Glu Asn Met Ala Gly Val 805 810 815 Phe Met Leu Val Ala Gly Gly Ile Val Ala Gly Ile Phe Leu Ile Phe 820 825 830 Ile Glu Ile Ala Tyr Lys Arg His Lys Asp Ala Arg Arg Lys Gln Met 835 840 845 Gln Leu Ala Phe Ala Ala Val Asn Val Trp Arg Lys Asn Leu Gln Asp 850 855 860 Arg Lys Ser Gly Arg Ala Glu Pro Asp Pro Lys Lys Lys Ala Thr Phe 865 870 875 880 Arg Ala Ile Thr Ser Thr Leu Ala Ser Ser Phe Lys Arg Arg Arg Ser 885 890 895 Ser Lys Asp Thr Ser Thr Gly Gly Gly Arg Gly Ala Leu Gln Asn Gln 900 905 910 Lys Asp Thr Val Leu Pro Arg Arg Ala Ile Glu Arg Glu Glu Gly Gln 915 920 925 Leu Gln Leu Cys Ser Arg His Arg Glu Ser 930 935 <210> SEQ ID NO 15 <211> LENGTH: 1464 <212> TYPE: PRT <213> ORGANISM: Rattus sp. <400> SEQUENCE: 15 Met Gly Arg Leu Gly Tyr Trp Thr Leu Leu Val Leu Pro Ala Leu Leu 1 5 10 15 Val Trp Arg Asp Pro Ala Gln Asn Ala Ala Ala Glu Lys Gly Pro Pro 20 25 30 Ala Leu Asn Ile Ala Val Leu Leu Gly His Ser His Asp Val Thr Glu 35 40 45 Arg Glu Leu Arg Asn Leu Trp Gly Pro Glu Gln Ala Thr Gly Leu Pro 50 55 60 Leu Asp Val Asn Val Val Ala Leu Leu Met Asn Arg Thr Asp Pro Lys 65 70 75 80 Ser Leu Ile Thr His Val Cys Asp Leu Met Ser Gly Ala Arg Ile His 85 90 95 Gly Leu Val Phe Gly Asp Asp Thr Asp Gln Glu Ala Val Ala Gln Met 100 105 110 Leu Asp Phe Ile Ser Ser Gln Thr Phe Ile Pro Ile Leu Gly Ile His 115 120 125 Gly Gly Ala Ser Met Ile Met Ala Asp Lys Asp Pro Thr Ser Thr Phe 130 135 140 Phe Gln Phe Gly Ala Ser Ile Gln Gln Gln Ala Thr Val Met Leu Lys 145 150 155 160 Ile Met Gln Asp Tyr Asp Trp His Val Phe Ser Leu Val Thr Thr Ile 165 170 175 Phe Pro Gly Tyr Arg Asp Phe Ile Ser Phe Ile Lys Thr Thr Val Asp 180 185 190 Asn Ser Phe Val Gly Trp Asp Met Gln Asn Val Ile Thr Leu Asp Thr 195 200 205 Ser Phe Glu Asp Ala Lys Thr Gln Val Gln Leu Lys Lys Ile His Ser 210 215 220 Ser Val Ile Leu Leu Tyr Cys Ser Lys Asp Glu Ala Val Leu Ile Leu 225 230 235 240 Ser Glu Ala Arg Ser Leu Gly Leu Thr Gly Tyr Asp Phe Phe Trp Ile 245 250 255 Val Pro Ser Leu Val Ser Gly Asn Thr Glu Leu Ile Pro Lys Glu Phe 260 265 270 Pro Ser Gly Leu Ile Ser Val Ser Tyr Asp Asp Trp Asp Tyr Ser Leu 275 280 285 Glu Ala Arg Val Arg Asp Gly Leu Gly Ile Leu Thr Thr Ala Ala Ser 290 295 300 Ser Met Leu Glu Lys Phe Ser Tyr Ile Pro Glu Ala Lys Ala Ser Cys 305 310 315 320 Tyr Gly Gln Ala Glu Lys Pro Glu Thr Pro Leu His Thr Leu His Gln 325 330 335 Phe Met Val Asn Val Thr Trp Asp Gly Lys Asp Leu Ser Phe Thr Glu 340 345 350 Glu Gly Tyr Gln Val His Pro Arg Leu Val Val Ile Val Leu Asn Lys 355 360 365 Asp Arg Glu Trp Glu Lys Val Gly Lys Trp Glu Asn Gln Thr Leu Ser 370 375 380 Leu Arg His Ala Val Trp Pro Arg Tyr Lys Ser Phe Ser Asp Cys Glu 385 390 395 400 Pro Asp Asp Asn His Leu Ser Ile Val Thr Leu Glu Glu Ala Pro Phe 405 410 415 Val Ile Val Glu Asp Ile Asp Pro Leu Thr Glu Thr Cys Val Arg Asn 420 425 430 Thr Val Pro Cys Arg Lys Phe Val Lys Ile Asn Asn Ser Thr Asn Glu 435 440 445 Gly Met Asn Val Lys Lys Cys Cys Lys Gly Phe Cys Ile Asp Ile Leu 450 455 460 Lys Lys Leu Ser Arg Thr Val Lys Phe Thr Tyr Asp Leu Tyr Leu Val 465 470 475 480 Thr Asn Gly Lys His Gly Lys Lys Val Asn Asn Val Trp Asn Gly Met 485 490 495 Ile Gly Glu Val Val Tyr Gln Arg Ala Val Met Ala Val Gly Ser Leu 500 505 510 Thr Ile Asn Glu Glu Arg Ser Glu Val Val Asp Phe Ser Val Pro Phe 515 520 525 Val Glu Thr Gly Ile Ser Val Met Val Ser Arg Ser Asn Gly Thr Val 530 535 540 Ser Pro Ser Ala Phe Leu Glu Pro Phe Ser Ala Ser Val Trp Val Met 545 550 555 560 Met Phe Val Met Leu Leu Ile Val Ser Ala Ile Ala Val Phe Val Phe 565 570 575 Glu Tyr Phe Ser Pro Val Gly Tyr Asn Arg Asn Leu Ala Lys Gly Lys 580 585 590 Ala Pro His Gly Pro Ser Phe Thr Ile Gly Lys Ala Ile Trp Leu Leu 595 600 605 Trp Gly Leu Val Phe Asn Asn Ser Val Pro Val Gln Asn Pro Lys Gly 610 615 620 Thr Thr Ser Lys Ile Met Val Ser Val Trp Ala Phe Phe Ala Val Ile 625 630 635 640 Phe Leu Ala Ser Tyr Thr Ala Asn Leu Ala Ala Phe Met Ile Gln Glu 645 650 655 Glu Phe Val Asp Gln Val Thr Gly Leu Ser Asp Lys Lys Phe Gln Arg 660 665 670 Pro His Asp Tyr Ser Pro Pro Phe Arg Phe Gly Thr Val Pro Asn Gly 675 680 685 Ser Thr Glu Arg Asn Ile Arg Asn Asn Tyr Pro Tyr Met His Gln Tyr 690 695 700 Met Thr Arg Phe Asn Gln Arg Gly Val Glu Asp Ala Leu Val Ser Leu 705 710 715 720 Lys Thr Gly Lys Leu Asp Ala Phe Ile Tyr Asp Ala Ala Val Leu Asn 725 730 735 Tyr Lys Ala Gly Arg Asp Glu Gly Cys Lys Leu Val Thr Ile Gly Ser 740 745 750 Gly Tyr Ile Phe Ala Thr Thr Gly Tyr Gly Ile Ala Leu Gln Lys Gly 755 760 765 Ser Pro Trp Lys Arg Gln Ile Asp Leu Ala Leu Leu Gln Phe Val Gly 770 775 780 Asp Gly Glu Met Glu Glu Leu Glu Thr Leu Trp Leu Thr Gly Ile Cys 785 790 795 800 His Asn Glu Lys Asn Glu Val Met Ser Ser Gln Leu Asp Ile Asp Asn 805 810 815 Met Ala Gly Val Phe Tyr Met Leu Ala Ala Ala Met Ala Leu Ser Leu 820 825 830 Ile Thr Phe Ile Trp Glu His Leu Phe Tyr Trp Lys Leu Arg Phe Cys 835 840 845 Phe Thr Gly Val Cys Ser Asp Arg Pro Gly Leu Leu Phe Ser Ile Ser 850 855 860 Arg Gly Ile Tyr Ser Cys Ile His Gly Val His Ile Glu Glu Lys Lys 865 870 875 880 Lys Ser Pro Asp Phe Asn Leu Thr Gly Ser Gln Ser Asn Met Leu Lys 885 890 895 Leu Leu Arg Ser Ala Lys Asn Ile Ser Asn Met Ser Asn Met Asn Ser 900 905 910 Ser Arg Met Asp Ser Pro Lys Arg Ala Thr Asp Phe Ile Gln Arg Gly 915 920 925 Ser Leu Ile Val Asp Met Val Ser Asp Lys Gly Asn Leu Ile Tyr Ser 930 935 940 Asp Asn Arg Ser Phe Gln Gly Lys Asp Ser Ile Phe Gly Asp Asn Met 945 950 955 960 Asn Glu Leu Gln Thr Phe Val Ala Asn Arg His Lys Asp Asn Leu Ser 965 970 975 Asn Tyr Val Phe Gln Gly Gln His Pro Leu Thr Leu Asn Asp Ser Asn 980 985 990 Pro Asn Thr Val Glu Val Ala Val Ser Thr Glu Ser Lys Gly Asn Ser 995 1000 1005 Arg Pro Arg Gln Leu Trp Lys Lys Ser Met Glu Ser Leu Arg Gln Asp 1010 1015 1020 Ser Leu Asn Gln Asn Pro Val Ser Gln Arg Asp Glu Lys Thr Ala Glu 1025 1030 1035 1040 Asn Arg Thr His Ser Leu Lys Ser Pro Arg Tyr Leu Pro Glu Glu Val 1045 1050 1055 Ala His Ser Asp Ile Ser Glu Thr Ser Ser Arg Ala Thr Cys His Arg 1060 1065 1070 Glu Pro Asp Asn Asn Lys Asn His Lys Thr Lys Asp Asn Phe Lys Arg 1075 1080 1085 Ser Met Ala Ser Lys Tyr Pro Lys Asp Cys Ser Asp Val Asp Arg Thr 1090 1095 1100 Tyr Met Lys Thr Lys Ala Ser Ser Pro Arg Asp Lys Ile Tyr Thr Ile 1105 1110 1115 1120 Asp Gly Glu Lys Glu Pro Ser Phe His Leu Asp Pro Pro Gln Phe Val 1125 1130 1135 Glu Asn Ile Thr Leu Pro Glu Asn Val Gly Phe Pro Asp Thr Tyr Gln 1140 1145 1150 Asp His Asn Glu Asn Phe Arg Lys Gly Asp Ser Thr Leu Pro Met Asn 1155 1160 1165 Arg Asn Pro Leu His Asn Glu Asp Gly Leu Pro Asn Asn Asp Gln Tyr 1170 1175 1180 Lys Leu Tyr Ala Lys His Phe Thr Leu Lys Asp Lys Gly Ser Pro His 1185 1190 1195 1200 Ser Glu Gly Ser Asp Arg Tyr Arg Gln Asn Ser Thr His Cys Arg Ser 1205 1210 1215 Cys Leu Ser Asn Leu Pro Thr Tyr Ser Gly His Phe Thr Met Arg Ser 1220 1225 1230 Pro Phe Lys Cys Asp Ala Cys Leu Arg Met Gly Asn Leu Tyr Asp Ile 1235 1240 1245 Asp Glu Asp Gln Met Leu Gln Glu Thr Gly Asn Pro Ala Thr Arg Glu 1250 1255 1260 Glu Val Tyr Gln Gln Asp Trp Ser Gln Asn Asn Ala Leu Gln Phe Gln 1265 1270 1275 1280 Lys Asn Lys Leu Arg Ile Asn Arg Gln His Ser Tyr Asp Asn Ile Leu 1285 1290 1295 Asp Lys Pro Arg Glu Ile Asp Leu Ser Arg Pro Ser Arg Ser Ile Ser 1300 1305 1310 Leu Lys Asp Arg Glu Arg Leu Leu Glu Gly Asn Leu Tyr Gly Ser Leu 1315 1320 1325 Phe Ser Val Pro Ser Ser Lys Leu Leu Gly Asn Lys Ser Ser Leu Phe 1330 1335 1340 Pro Gln Gly Leu Glu Asp Ser Lys Arg Ser Lys Ser Leu Leu Pro Asp 1345 1350 1355 1360 His Ala Ser Asp Asn Pro Phe Leu His Thr Tyr Gly Asp Asp Gln Arg 1365 1370 1375 Leu Val Ile Gly Arg Cys Pro Ser Asp Pro Tyr Lys His Ser Leu Pro 1380 1385 1390 Ser Gln Ala Val Asn Asp Ser Tyr Leu Arg Ser Ser Leu Arg Ser Thr 1395 1400 1405 Ala Ser Tyr Cys Ser Arg Asp Ser Arg Gly His Ser Asp Val Tyr Ile 1410 1415 1420 Ser Glu His Val Met Pro Tyr Ala Ala Asn Lys Asn Thr Met Tyr Ser 1425 1430 1435 1440 Thr Pro Arg Val Leu Asn Ser Cys Ser Asn Arg Arg Val Tyr Lys Lys 1445 1450 1455 Met Pro Ser Ile Glu Ser Asp Val 1460 <210> SEQ ID NO 16 <211> LENGTH: 1115 <212> TYPE: PRT <213> ORGANISM: Rattus sp. <400> SEQUENCE: 16 Met Arg Arg Leu Ser Leu Trp Trp Leu Leu Ser Arg Val Cys Leu Leu 1 5 10 15 Leu Pro Pro Pro Cys Ala Leu Val Leu Ala Gly Val Pro Ser Ser Ser 20 25 30 Ser His Pro Gln Pro Cys Gln Ile Leu Lys Arg Ile Gly His Ala Val 35 40 45 Arg Val Gly Ala Val His Leu Gln Pro Trp Thr Thr Ala Pro Arg Ala 50 55 60 Ala Ser Arg Ala Gln Glu Gly Gly Arg Ala Gly Ala Gln Arg Asp Asp 65 70 75 80 Pro Glu Ser Gly Thr Trp Arg Pro Pro Ala Pro Ser Gln Gly Ala Arg 85 90 95 Trp Leu Gly Ser Ala Leu His Gly Arg Gly Pro Pro Gly Ser Arg Lys 100 105 110 Leu Gly Glu Gly Ala Gly Ala Glu Thr Leu Trp Pro Arg Asp Ala Leu 115 120 125 Leu Phe Ala Val Glu Asn Leu Asn Arg Val Glu Gly Leu Leu Pro Tyr 130 135 140 Asn Leu Ser Leu Glu Val Val Met Ala Ile Glu Ala Gly Leu Gly Asp 145 150 155 160 Leu Pro Leu Met Pro Phe Ser Ser Pro Ser Ser Pro Trp Ser Ser Asp 165 170 175 Pro Phe Ser Phe Leu Gln Ser Val Cys His Thr Val Val Val Gln Gly 180 185 190 Val Ser Ala Leu Leu Ala Phe Pro Gln Ser Gln Gly Glu Met Met Glu 195 200 205 Leu Asp Leu Val Ser Ser Val Leu His Ile Pro Val Leu Ser Ile Val 210 215 220 Arg His Glu Phe Pro Arg Glu Ser Gln Asn Pro Leu His Leu Gln Leu 225 230 235 240 Ser Leu Glu Asn Ser Leu Ser Ser Asp Ala Asp Val Thr Val Ser Ile 245 250 255 Leu Thr Met Asn Asn Trp Tyr Asn Phe Ser Leu Leu Leu Cys Gln Glu 260 265 270 Asp Trp Asn Ile Thr Asp Phe Leu Leu Leu Thr Glu Asn Asn Ser Lys 275 280 285 Phe His Leu Glu Ser Val Ile Asn Ile Thr Ala Asn Leu Ser Ser Thr 290 295 300 Lys Asp Leu Leu Ser Phe Leu Gln Val Gln Met Asp Asn Ile Arg Asn 305 310 315 320 Ser Thr Pro Thr Met Val Met Phe Gly Cys Asp Met Asp Ser Ile Arg 325 330 335 Gln Ile Phe Glu Met Ser Thr Gln Phe Gly Leu Ser Pro Pro Glu Leu 340 345 350 His Trp Val Leu Gly Asp Ser Gln Asn Val Glu Glu Leu Arg Thr Glu 355 360 365 Gly Leu Pro Leu Gly Leu Ile Ala His Gly Lys Thr Thr Gln Ser Val 370 375 380 Phe Glu Tyr Tyr Val Gln Asp Ala Met Glu Leu Val Ala Arg Ala Val 385 390 395 400 Ala Thr Ala Thr Met Ile Gln Pro Glu Leu Ala Leu Leu Pro Ser Thr 405 410 415 Met Asn Cys Met Asp Val Lys Thr Thr Asn Leu Thr Ser Gly Gln Tyr 420 425 430 Leu Ser Arg Phe Leu Ala Asn Thr Thr Phe Arg Gly Leu Ser Gly Ser 435 440 445 Ile Lys Val Lys Gly Ser Thr Ile Ile Ser Ser Glu Asn Asn Phe Phe 450 455 460 Ile Trp Asn Leu Gln His Asp Pro Met Gly Lys Pro Met Trp Thr Arg 465 470 475 480 Leu Gly Ser Trp Gln Gly Gly Arg Ile Val Met Asp Ser Gly Ile Trp 485 490 495 Pro Glu Gln Ala Gln Arg His Lys Thr His Phe Gln His Pro Asn Lys 500 505 510 Leu His Leu Arg Val Val Thr Leu Ile Glu His Pro Phe Val Phe Thr 515 520 525 Arg Glu Val Asp Asp Glu Gly Leu Cys Pro Ala Gly Gln Leu Cys Leu 530 535 540 Asp Pro Met Thr Asn Asp Ser Ser Met Leu Asp Arg Leu Phe Ser Ser 545 550 555 560 Leu His Ser Ser Asn Asp Thr Val Pro Ile Lys Phe Lys Lys Cys Cys 565 570 575 Tyr Gly Tyr Cys Ile Asp Leu Leu Glu Gln Leu Ala Glu Asp Met Asn 580 585 590 Phe Asp Phe Asp Leu Tyr Ile Val Gly Asp Gly Lys Tyr Gly Ala Trp 595 600 605 Lys Asn Gly His Trp Thr Gly Leu Val Gly Asp Leu Leu Ser Gly Thr 610 615 620 Ala Asn Met Ala Val Thr Ser Phe Ser Ile Asn Thr Ala Arg Ser Gln 625 630 635 640 Val Ile Asp Phe Thr Ser Pro Phe Phe Ser Thr Ser Leu Gly Ile Leu 645 650 655 Val Arg Thr Arg Asp Thr Ala Ala Pro Ile Gly Ala Phe Met Trp Pro 660 665 670 Leu His Trp Thr Met Trp Leu Gly Ile Phe Val Ala Leu His Ile Thr 675 680 685 Ala Ile Phe Leu Thr Leu Tyr Glu Trp Lys Ser Pro Phe Gly Met Thr 690 695 700 Pro Lys Gly Arg Asn Arg Asn Lys Val Phe Ser Phe Ser Ser Ala Leu 705 710 715 720 Asn Val Cys Tyr Ala Leu Leu Phe Gly Arg Thr Ala Ala Ile Lys Pro 725 730 735 Pro Lys Cys Trp Thr Gly Arg Phe Leu Met Asn Leu Trp Ala Ile Phe 740 745 750 Cys Met Phe Cys Leu Ser Thr Tyr Thr Ala Asn Leu Ala Ala Val Met 755 760 765 Val Gly Glu Lys Ile Tyr Glu Glu Leu Ser Gly Ile His Asp Pro Lys 770 775 780 Leu His His Pro Ser Gln Gly Phe Arg Phe Gly Thr Val Arg Glu Ser 785 790 795 800 Ser Ala Glu Asp Tyr Val Arg Gln Ser Phe Pro Glu Met His Glu Tyr 805 810 815 Met Arg Arg Tyr Asn Val Pro Ala Thr Pro Asp Gly Val Gln Tyr Leu 820 825 830 Lys Asn Asp Pro Glu Lys Leu Asp Ala Phe Ile Met Asp Lys Ala Leu 835 840 845 Leu Asp Tyr Glu Val Ser Ile Asp Ala Asp Cys Lys Leu Leu Thr Val 850 855 860 Gly Lys Pro Phe Ala Ile Glu Gly Tyr Gly Ile Gly Leu Pro Pro Asn 865 870 875 880 Ser Pro Leu Thr Ser Asn Ile Ser Glu Leu Ile Ser Gln Tyr Lys Ser 885 890 895 His Gly Phe Met Asp Val Leu His Asp Lys Trp Tyr Lys Val Val Pro 900 905 910 Cys Gly Lys Arg Ser Phe Ala Val Thr Glu Thr Leu Gln Met Gly Ile 915 920 925 Lys His Phe Ser Gly Leu Phe Val Leu Leu Cys Ile Gly Phe Gly Leu 930 935 940 Ser Ile Leu Thr Thr Ile Gly Glu His Ile Val His Arg Leu Leu Leu 945 950 955 960 Pro Arg Ile Lys Asn Lys Ser Lys Leu Gln Tyr Trp Leu His Thr Ser 965 970 975 Gln Arg Phe His Arg Ala Leu Asn Thr Ser Phe Val Glu Glu Lys Gln 980 985 990 Pro Arg Ser Lys Thr Lys Arg Val Glu Lys Arg Ser Asn Leu Gly Pro 995 1000 1005 Gln Gln Leu Met Val Trp Asn Thr Ser Asn Leu Ser His Asp Asn Gln 1010 1015 1020 Arg Lys Tyr Ile Phe Asn Asp Glu Glu Gly Gln Asn Gln Leu Gly Thr 1025 1030 1035 1040 Gln Ala His Gln Asp Ile Pro Leu Pro Gln Arg Arg Arg Glu Leu Pro 1045 1050 1055 Ala Ser Leu Thr Thr Asn Gly Lys Ala Asp Ser Leu Asn Val Thr Arg 1060 1065 1070 Ser Ser Val Ile Gln Glu Leu Ser Glu Leu Glu Lys Gln Ile Gln Val 1075 1080 1085 Ile Arg Gln Glu Leu Gln Leu Ala Val Ser Arg Lys Thr Glu Leu Glu 1090 1095 1100 Glu Tyr Gln Lys Thr Asn Arg Thr Cys Glu Ser 1105 1110 1115 <210> SEQ ID NO 17 <211> LENGTH: 21 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: synthetic oligonucleotide <400> SEQUENCE: 17 tgctgctatg gctactgcat c 21 <210> SEQ ID NO 18 <211> LENGTH: 23 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: synthetic oligonucleotide <400> SEQUENCE: 18 atgacagcag ccaggttggc cgt 23 <210> SEQ ID NO 19 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: synthetic oligonucleotide <400> SEQUENCE: 19 cacacatggc tgtgaccagc 20 <210> SEQ ID NO 20 <211> LENGTH: 21 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: synthetic oligonucleotide <400> SEQUENCE: 20 agaatggcat agcacaggtt g 21 <210> SEQ ID NO 21 <211> LENGTH: 6 <212> TYPE: PRT <213> ORGANISM: Rattus sp. <400> SEQUENCE: 21 Ala Asp Gly Lys Phe Gly 1 5 <210> SEQ ID NO 22 <211> LENGTH: 6 <212> TYPE: PRT <213> ORGANISM: Rattus sp. <400> SEQUENCE: 22 Thr Asn Gly Lys His Gly 1 5 <210> SEQ ID NO 23 <211> LENGTH: 6 <212> TYPE: PRT <213> ORGANISM: Rattus sp. <400> SEQUENCE: 23 Thr Asn Gly Lys His Gly 1 5 <210> SEQ ID NO 24 <211> LENGTH: 6 <212> TYPE: PRT <213> ORGANISM: Rattus sp. <400> SEQUENCE: 24 Thr Asn Gly Lys His Gly 1 5 <210> SEQ ID NO 25 <211> LENGTH: 6 <212> TYPE: PRT <213> ORGANISM: Rattus sp. <400> SEQUENCE: 25 Thr Asn Gly Lys His Gly 1 5 <210> SEQ ID NO 26 <211> LENGTH: 6 <212> TYPE: PRT <213> ORGANISM: Rattus sp. <400> SEQUENCE: 26 Gly Asp Gly Lys Tyr Gly 1 5 <210> SEQ ID NO 27 <211> LENGTH: 6 <212> TYPE: PRT <213> ORGANISM: Rattus sp. <400> SEQUENCE: 27 Gly Asp Gly Lys Tyr Gly 1 5 <210> SEQ ID NO 28 <211> LENGTH: 6 <212> TYPE: PRT <213> ORGANISM: Rattus sp. <400> SEQUENCE: 28 Gly Asp Gly Lys Tyr Gly 1 5 <210> SEQ ID NO 29 <211> LENGTH: 11 <212> TYPE: PRT <213> ORGANISM: Rattus sp. <400> SEQUENCE: 29 Ala Pro Leu Thr Ile Asn Asn Glu Arg Ala Gln 1 5 10 <210> SEQ ID NO 30 <211> LENGTH: 11 <212> TYPE: PRT <213> ORGANISM: Rattus sp. <400> SEQUENCE: 30 Gly Ser Leu Thr Ile Asn Glu Glu Arg Ser Glu 1 5 10 <210> SEQ ID NO 31 <211> LENGTH: 11 <212> TYPE: PRT <213> ORGANISM: Rattus sp. <400> SEQUENCE: 31 Gly Ser Leu Thr Ile Asn Glu Glu Arg Ser Glu 1 5 10 <210> SEQ ID NO 32 <211> LENGTH: 11 <212> TYPE: PRT <213> ORGANISM: Rattus sp. <400> SEQUENCE: 32 Gly Ser Leu Thr Ile Asn Glu Glu Arg Ser Glu 1 5 10 <210> SEQ ID NO 33 <211> LENGTH: 11 <212> TYPE: PRT <213> ORGANISM: Rattus sp. <400> SEQUENCE: 33 Gly Ser Leu Thr Ile Asn Glu Glu Arg Ser Glu 1 5 10 <210> SEQ ID NO 34 <211> LENGTH: 11 <212> TYPE: PRT <213> ORGANISM: Rattus sp. <400> SEQUENCE: 34 Thr Ser Phe Ser Ile Asn Thr Ala Arg Ser Gln 1 5 10 <210> SEQ ID NO 35 <211> LENGTH: 11 <212> TYPE: PRT <213> ORGANISM: Rattus sp. <400> SEQUENCE: 35 Thr Ser Phe Ser Ile Asn Ser Ala Arg Ser Gln 1 5 10 <210> SEQ ID NO 36 <211> LENGTH: 11 <212> TYPE: PRT <213> ORGANISM: Rattus sp. <400> SEQUENCE: 36 Ala Pro Leu Thr Ile Thr Leu Val Arg Glu Glu 1 5 10 <210> SEQ ID NO 37 <211> LENGTH: 9 <212> TYPE: PRT <213> ORGANISM: Rattus sp. <400> SEQUENCE: 37 Ala Thr Val Lys Gln Ser Ser Val Asp 1 5 <210> SEQ ID NO 38 <211> LENGTH: 9 <212> TYPE: PRT <213> ORGANISM: Rattus sp. <400> SEQUENCE: 38 Gly Thr Val Pro Asn Gly Ser Thr Glu 1 5 <210> SEQ ID NO 39 <211> LENGTH: 9 <212> TYPE: PRT <213> ORGANISM: Rattus sp. <400> SEQUENCE: 39 Gly Thr Val Pro Asn Gly Ser Thr Glu 1 5 <210> SEQ ID NO 40 <211> LENGTH: 9 <212> TYPE: PRT <213> ORGANISM: Rattus sp. <400> SEQUENCE: 40 Gly Thr Val Pro Asn Gly Ser Thr Glu 1 5 <210> SEQ ID NO 41 <211> LENGTH: 9 <212> TYPE: PRT <213> ORGANISM: Rattus sp. <400> SEQUENCE: 41 Gly Thr Val Pro Asn Gly Ser Thr Glu 1 5 <210> SEQ ID NO 42 <211> LENGTH: 9 <212> TYPE: PRT <213> ORGANISM: Rattus sp. <400> SEQUENCE: 42 Gly Thr Val Arg Glu Ser Ser Ala Glu 1 5 <210> SEQ ID NO 43 <211> LENGTH: 9 <212> TYPE: PRT <213> ORGANISM: Rattus sp. <400> SEQUENCE: 43 Gly Thr Val Trp Glu Ser Ser Ala Glu 1 5 <210> SEQ ID NO 44 <211> LENGTH: 9 <212> TYPE: PRT <213> ORGANISM: Rattus sp. <400> SEQUENCE: 44 Gly Thr Leu Asp Ser Gly Ser Thr Lys 1 5 <210> SEQ ID NO 45 <211> LENGTH: 25 <212> TYPE: PRT <213> ORGANISM: Rattus sp. <400> SEQUENCE: 45 Asp Ala Leu Thr Leu Ser Ser Ala Met Trp Phe Ser Trp Gly Val Leu 1 5 10 15 Leu Asn Ser Gly Ile Gly Glu Gly Ala 20 25 <210> SEQ ID NO 46 <211> LENGTH: 25 <212> TYPE: PRT <213> ORGANISM: Rattus sp. <400> SEQUENCE: 46 Pro Ser Phe Thr Ile Gly Lys Ala Ile Trp Leu Leu Trp Gly Leu Val 1 5 10 15 Phe Asn Asn Ser Val Pro Val Gln Asn 20 25 <210> SEQ ID NO 47 <211> LENGTH: 25 <212> TYPE: PRT <213> ORGANISM: Rattus sp. <400> SEQUENCE: 47 Pro Ser Phe Thr Ile Gly Lys Ala Ile Trp Leu Leu Trp Gly Leu Val 1 5 10 15 Phe Asn Asn Ser Val Pro Val Gln Asn 20 25 <210> SEQ ID NO 48 <211> LENGTH: 25 <212> TYPE: PRT <213> ORGANISM: Rattus sp. <400> SEQUENCE: 48 Pro Ser Phe Thr Ile Gly Lys Ser Val Trp Leu Leu Trp Ala Leu Val 1 5 10 15 Phe Asn Asn Ser Val Pro Ile Glu Asn 20 25 <210> SEQ ID NO 49 <211> LENGTH: 25 <212> TYPE: PRT <213> ORGANISM: Rattus sp. <400> SEQUENCE: 49 Ser Thr Phe Thr Ile Gly Lys Ser Ile Trp Leu Leu Trp Ala Leu Val 1 5 10 15 Phe Asn Asn Ser Val Pro Val Glu Asn 20 25 <210> SEQ ID NO 50 <211> LENGTH: 25 <212> TYPE: PRT <213> ORGANISM: Rattus sp. <400> SEQUENCE: 50 Lys Val Phe Ser Phe Ser Ser Ala Leu Asn Val Cys Tyr Ala Leu Leu 1 5 10 15 Phe Gly Arg Thr Ala Ala Ile Lys Pro 20 25 <210> SEQ ID NO 51 <211> LENGTH: 25 <212> TYPE: PRT <213> ORGANISM: Rattus sp. <400> SEQUENCE: 51 Thr Val Phe Ser Tyr Ser Ser Ala Leu Asn Leu Cys Tyr Ala Ile Leu 1 5 10 15 Phe Gly Arg Thr Val Ser Ser Lys Thr 20 25 <210> SEQ ID NO 52 <211> LENGTH: 30 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: syntheitc oligonucleotide <400> SEQUENCE: 52 ctagcggcgc gccttaatta agtttaaacg 30 <210> SEQ ID NO 53 <211> LENGTH: 30 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: syntheitc oligonucleotide <400> SEQUENCE: 53 ctagcgttta aacttaatta aggcgcgccg 30 

What is claimed is:
 1. An isolated NR3B nucleic acid molecule, comprising a nucleotide sequence which encodes a NR3B polypeptide having at least 80% identity to the amino acid sequence designated SEQ ID NO:6, wherein said polypeptide does not consist of the amino acid sequence designated SEQ ID NO:10 or
 12. 2. The isolated nucleic acid molecule of claim 1, wherein said NR3B polypeptide comprises the amino acid sequence designated SEQ ID NO:6.
 3. The isolated nucleic acid molecule of claim 1, wherein said NR3B polypeptide comprises the amino acid sequence designated SEQ ID NO:4.
 4. The isolated nucleic acid molecule of claim 1, wherein said NR3B polypeptide comprises the amino acid sequence designated SEQ ID NO:2.
 5. The isolated nucleic acid molecule of claim 1, wherein said NR3B polypeptide comprises the amino acid sequence designated SEQ ID NO:8.
 6. The isolated nucleic acid molecule of claim 1, operatively linked to a promoter of gene expression.
 7. A vector comprising the isolated nucleic acid molecule of claim
 1. 8. A cell comprising the isolated nucleic acid molecule of claim
 1. 9. The cell of claim 8, wherein said NR3B polypeptide is expressed at the cell membrane.
 10. The cell of claim 8, wherein an NR1 polypeptide is further expressed at the cell membrane.
 11. The cell of claim 10, wherein said NR1 polypeptide comprises a human, rat or mouse NR1 amino acid sequence.
 12. The cell of claim 10, wherein said NR1 polypeptide comprises the amino acid sequence designated SEQ ID NO:14.
 13. A method of producing an NR3B polypeptide, comprising expressing the nucleic acid molecule of claim 1 in vitro or in a cell under conditions suitable for expression of said polypeptide.
 14. An isolated NR3B nucleic acid molecule, comprising a nucleotide sequence which encodes a functional fragment of an NR3B polypeptide, wherein said NR3B polypeptide comprises an amino acid sequence designated SEQ ID NO:2, 4, 6 or 8, wherein said functional fragment does not consist of the amino acid sequence designated SEQ ID NO:10 or
 12. 15. The isolated nucleic acid molecule of claim 14, wherein said functional fragment binds glycine.
 16. An isolated NR3B polynucleotide, comprising at least 17 contiguous nucleotides from the nucleotide sequence designated SEQ ID NO:1, 3, 5 or 7, or from the complement thereof, wherein said nucleic acid molecule does not consist of the nucleotide sequence designated SEQ ID NO:9, 11 or
 13. 17. A method for detecting a nucleic acid molecule encoding a NR3B polypeptide in a sample, comprising contacting said sample with one or more polynucleotides according to claim 16, and detecting specific hybridization to said polynucleotide, thereby detecting a nucleic acid molecule encoding an NR3B polypeptide in said sample.
 18. An isolated NR3B polypeptide, comprising an amino acid sequence having at least 80% identity to SEQ ID NO:6, wherein said polypeptide does not consist of the amino acid sequence designated SEQ ID NO:10 or
 12. 19. The isolated polypeptide of claim 18, wherein said polypeptide comprises the amino acid sequence designated SEQ ID NO:6.
 20. The isolated polypeptide of claim 18, wherein said polypeptide comprises the amino acid sequence designated SEQ ID NO:4.
 21. The isolated polypeptide of claim 18, wherein said polypeptide comprises the amino acid sequence designated SEQ ID NO:2.
 22. The isolated polypeptide of claim 18, wherein said polypeptide comprises the amino acid sequence designated SEQ ID NO:8.
 23. The isolated polypeptide of claim 18, further comprising a membrane.
 24. The isolated polypeptide of claim 18, which is associated with an NR1 polypeptide to form an excitatory glycine receptor.
 25. The isolated polypeptide of claim 18, which is bound to glycine.
 26. A functional fragment of an NR3B polypeptide, wherein said NR3B polypeptide comprises an amino acid sequence designated SEQ ID NO:2, 4, 6 or 8, wherein said functional fragment does not consist of the amino acid sequence designated SEQ ID NO:10 or
 12. 27. The isolated polypeptide of claim 26, wherein said functional fragment binds glycine.
 28. An isolated NR3B peptide, comprising at least 8 contiguous residues of the amino acid sequence designated SEQ ID NO:2, 4, 6 or 8, wherein said peptide does not consist of the amino acid sequence designated SEQ ID NO:10 or
 12. 29. An isolated antibody or antigen binding fragment thereof which specifically binds the isolated NR3B polypeptide of claim
 18. 30. A method of detecting an NR3B polypeptide in a sample, comprising contacting said sample with the antibody of claim 29, and detecting the presence of specific binding of said antibody to said sample, thereby detecting an NR3B polypeptide in said sample.
 31. A method of detecting an NR3B ligand, comprising: (a) contacting the polypeptide of claim 18 with one or more candidate compounds under conditions suitable for detecting binding to said polypeptide; and (b) detecting a candidate compound that binds said polypeptide, wherein said compound is characterized as an NR3B ligand.
 32. A method of detecting an NR3B ligand, comprising: (a) contacting the functional fragment of claim 27 with one or more candidate compounds under conditions suitable for detecting binding to said fragment; and (b) detecting a candidate compound that binds said fragment, wherein said compound is characterized as an NR3B ligand.
 33. A composition, comprising an isolated excitatory glycine receptor.
 34. The composition of claim 33, further comprising a cell membrane.
 35. The composition of claim 34, wherein said cell is a Xenopus oocyte or mammalian cell.
 36. The composition of claim 33, wherein said excitatory glycine receptor comprises an NR3B polypeptide and an NR1 polypeptide.
 37. The composition of claim 36, wherein said excitatory glycine receptor further comprises an NR2 polypeptide.
 38. The composition of claim 36, wherein said NR3B polypeptide comprises the amino acid sequence designated SEQ ID NO:2, 4, 6 or
 8. 39. The composition of claim 36, wherein said NR1 polypeptide comprises a naturally-occurring human, rat or mouse NR1 amino acid sequence.
 40. The composition of claim 33, wherein said excitatory glycine receptor comprises an NR3A polypeptide and an NR1 polypeptide.
 41. The composition of claim 40, wherein said excitatory glycine receptor further comprises an NR2 polypeptide.
 42. The composition of claim 40, wherein said NR3A polypeptide comprises a naturally-occurring human or rat NR3A amino acid sequence.
 43. The composition of claim 40, wherein said NR1 polypeptide comprises a naturally-occurring human, rat or mouse NR1 amino acid sequence.
 44. A method of detecting an excitatory glycine receptor ligand, comprising: (a) contacting an excitatory glycine receptor with one or more candidate compounds under conditions suitable for detecting binding to said receptor; and (b) detecting a candidate compound that binds said receptor, wherein said compound is characterized as an excitatory glycine receptor ligand.
 45. The method of claim 44, wherein said excitatory glycine receptor comprises an NR3B polypeptide and an NR1 polypeptide.
 46. The method of claim 45, wherein said excitatory glycine receptor further comprises an NR2 polypeptide.
 47. The method of claim 45, wherein said NR3B polypeptide comprises the amino acid sequence designated SEQ ID NO:2, 4, 6 or
 8. 48. The method of claim 45, wherein said NR1 polypeptide comprises a naturally-occurring human, rat or mouse NR1 amino acid sequence.
 49. The method of claim 44, wherein said excitatory glycine receptor comprises an NR3A polypeptide and an NR1 polypeptide.
 50. The method of claim 49, wherein said excitatory glycine receptor further comprises an NR2 polypeptide.
 51. The method of claim 49, wherein said NR3A polypeptide comprises a naturally-occurring human or rat NR3A amino acid sequence.
 52. The method of claim 49, wherein said NR1 polypeptide comprises a naturally-occurring human, rat or mouse NR1 amino acid sequence.
 53. The method of claim 44, wherein said contacting occurs in the presence of glycine.
 54. A method of detecting an excitatory glycine receptor agonist or antagonist, comprising: (a) contacting an excitatory glycine receptor with one or more candidate compounds under conditions suitable for detecting receptor activation; and (b) detecting a candidate compound that alters said receptor activation, wherein said compound is characterized as an excitatory glycine receptor agonist or antagonist.
 55. The method of claim 54, wherein said excitatory glycine receptor comprises an NR3B polypeptide and an NR1 polypeptide.
 56. The method of claim 55, wherein said excitatory glycine receptor further comprises an NR2 polypeptide.
 57. The method of claim 55, wherein said NR3B polypeptide comprises the amino acid sequence designated SEQ ID NO:2, 4, 6 or
 8. 58. The method of claim 55, wherein said NR1 polypeptide comprises a naturally-occurring human, rat or mouse NR1 amino acid sequence.
 59. The method of claim 54, wherein said excitatory glycine receptor comprises an NR3A polypeptide and an NR1 polypeptide.
 60. The method of claim 59, wherein said excitatory glycine receptor further comprises an NR2 polypeptide.
 61. The method of claim 59, wherein said NR3A polypeptide comprises a naturally-occurring human or rat NR3A amino acid sequence.
 62. The method of claim 59, wherein said NR1 polypeptide comprises a naturally-occurring human, rat or mouse NR1 amino acid sequence.
 63. The method of claim 54, wherein said receptor activation is detected by assaying whole-cell currents.
 64. The method of claim 54, wherein said receptor activation is detected by assaying single-channel currents.
 65. The method of claim 54, wherein said receptor activation is detected by assaying ion fluxes using ion indicators.
 66. The method of claim 63, wherein said cell is a Xenopus oocyte or mammalian cell that expresses said excitatory glycine receptor.
 67. The method of claim 54, wherein said contacting occurs in the presence of glycine.
 68. A method of modulating a cellular response to glycine or glutamate, comprising: (a) introducing the nucleic acid molecule of claim 1 into a cell; and (b) expressing the NR3B polypeptide encoded by said nucleic acid molecule in said cell, whereby expression of said polypeptide modulates a cellular response to glycine or glutamate.
 69. A method of modulating a cellular response to glycine or glutamate, comprising: (a) introducing the nucleic acid molecule of claim 1 into a cell; and (b) expressing the NR3B functional fragment encoded by said nucleic acid molecule in said cell, whereby expression of said functional fragment modulates a cellular response to glycine or glutamate.
 70. A method of modulating a cellular response to glycine or glutamate, comprising introducing an antisense nucleic acid molecule, a ribozyme molecule or a small interfering RNA (siRNA) molecule into said cell, wherein said molecule hybridizes to any of SEQ ID NO:1, 3, 5 or 7 and prevents translation of the encoded NR3B polypeptide. 